Virus detection method, virus detection device, virus determination program, pressure determination method, and pressure determination device
阅读说明：本技术 病毒检测方法、病毒检测装置、病毒判定程序、压力判定方法以及压力判定装置 (Virus detection method, virus detection device, virus determination program, pressure determination method, and pressure determination device ) 是由 西田幸二 桥田德康 谷口正辉 筒井真楠 于 2019-07-18 设计创作，主要内容包括：本发明提供一种病毒检测方法、病毒检测装置、病毒判定程序、压力判定方法以及压力判定装置。一种病毒检测方法,包括以下步骤：接触步骤,经由设置在隔壁上的通孔部而使包含被检者体液的液体样本和电解液接触；电流检测步骤,对于所述通孔部,向所述液体样本和所述电解液施加电压,获取流过通孔部的离子电流的波形；病毒判定步骤,基于波形判定体液所含的病毒的种类；所述病毒判定步骤是将所述波形和与预先获取的已知病毒对应的波形信息进行对比来判定病毒的种类。(The invention provides a virus detection method, a virus detection device, a virus determination program, a pressure determination method, and a pressure determination device. A method of detecting a virus comprising the steps of: a contact step of bringing a liquid sample containing a body fluid of a subject into contact with an electrolytic solution via a through hole portion provided in a partition wall; a current detection step of applying a voltage to the liquid sample and the electrolytic solution to the through hole portion to obtain a waveform of an ion current flowing through the through hole portion; a virus determination step of determining the type of virus contained in the body fluid based on the waveform; the virus determining step determines the type of the virus by comparing the waveform with waveform information corresponding to a known virus acquired in advance.)
1. A method for detecting a virus, comprising the steps of:
a contact step of bringing a liquid sample containing a body fluid of a subject into contact with an electrolytic solution via a through hole provided in a partition wall;
a current detection step of applying a voltage to the liquid sample and the electrolytic solution to the through-hole portion to obtain a waveform of an ion current flowing through the through-hole portion; and
a virus determination step of determining a virus type contained in the body fluid based on the waveform,
and a virus determination step of comparing the waveform with waveform information corresponding to a known virus acquired in advance to determine the type of the virus within 10 minutes.
2. The virus detection method according to claim 1, wherein the current detection step comprises: a passing step of passing the microparticles contained in the body fluid through the through-hole portion by electrophoresis.
3. The method according to claim 1 or 2, wherein the known virus is a herpes virus.
4. The method for detecting a virus according to any one of claims 1 to 3, wherein the body fluid is selected from the group consisting of tears and intraocular fluid.
5. The virus detection method according to any one of claims 1 to 4, wherein the current detection step comprises: a separation step of separating viruses contained in the body fluid and foreign substances other than viruses contained in the body fluid by electroosmotic flow.
6. The method according to any one of claims 1 to 5, wherein the disease is identified based on the type of the virus determined in the virus determination step.
7. A virus detection device, comprising:
a sample storage unit for storing a liquid sample containing a body fluid of a subject;
an electrolyte storage unit that stores an electrolyte;
a partition wall portion that partitions the sample storage portion and the electrolyte solution storage portion;
a through hole portion provided in the partition wall portion and communicating the sample storage portion and the electrolyte solution storage portion;
a detection unit that obtains a waveform of an ion current flowing through the through hole unit by applying a voltage to the liquid sample and the electrolyte;
a storage unit that stores a learned model that has been machine-learned so that, when the waveform is input, a virus type corresponding to waveform information of a known virus type is output; and
and a virus determination unit that applies the waveform acquired from the detection unit to the learned model read from the storage unit to determine a virus type contained in the body fluid.
8. A virus detection device, comprising:
a sample storage section that stores a liquid sample containing a body fluid of a subject selected from the group consisting of tears and intraocular fluid;
an electrolyte storage unit that stores an electrolyte;
a partition wall portion that partitions the sample storage portion and the electrolyte solution storage portion;
a through hole portion provided in the partition wall portion and having hydrophilicity for communicating the sample storage portion and the electrolyte storage portion;
a detection unit that obtains a waveform of an ion current flowing through the through hole unit by applying a voltage to the liquid sample and the electrolyte;
a storage unit for storing waveform information of a known virus type; and
a virus determination unit for determining a virus type contained in the body fluid based on the waveform,
the virus determination unit determines the type of virus contained in the body fluid by comparing the waveform acquired from the detection unit with the waveform information of the known type of virus read from the storage unit.
9. The virus detection device according to claim 7 or 8, wherein the liquid sample comprises glutathione.
10. The virus detection apparatus according to any one of claims 7 to 9, wherein the virus determination unit determines the type of the virus within 10 minutes, and is portable.
11. A virus determination program for causing a computer to execute a process of determining a virus type contained in a body fluid of a subject, comprising:
a reception process of receiving a waveform of an ion current obtained by applying a voltage to a liquid sample containing the body fluid and an electrolytic solution;
a virus determination process of determining a virus type contained in the body fluid based on the waveform received in the reception process; and
a transmission process of transmitting a judgment result in the virus judgment process,
the virus determination process determines the type of virus based on a learned model that has been machine-learned so that, when the waveform is input, a virus type corresponding to waveform information of a known virus type is output.
12. A pressure determination method, characterized by comprising the steps of:
a contact step of bringing a liquid sample containing a body fluid of a subject into contact with an electrolytic solution via a through hole portion provided in a partition wall;
a current detection step of applying a voltage to the liquid sample and the electrolytic solution to the through hole portion to obtain a waveform of an ion current flowing through the through hole portion; and
a determination step of comparing the waveform with specific pressure state information to determine the pressure state of the subject.
13. The pressure determination method according to claim 12, wherein the current detection step includes: a passing step of passing the microparticles contained in the body fluid through the through-hole portion by electrophoresis,
the determining step includes: a counting step of counting the number of the fine particles passing through the through hole portion based on the waveform; and a comparison step of comparing the number of the particles with the pressure state information;
the pressure state information includes information on a relationship between the number of the microparticles contained in the body fluid and the pressure intensity.
14. The method for pressure determination according to claim 13, wherein the microparticle is a herpesvirus.
15. The method according to any one of claims 12 to 14, wherein the body fluid is tear fluid.
16. A pressure determination device, characterized by comprising:
a sample storage unit for storing a liquid sample containing a body fluid of a subject;
an electrolyte storage unit that stores an electrolyte;
a partition wall portion that partitions the sample storage portion and the electrolyte solution storage portion;
a through hole portion provided in the partition wall portion and communicating the sample storage portion and the electrolyte solution storage portion;
a detection unit that obtains a waveform of an ion current flowing through the through hole unit by applying a voltage to the liquid sample and the electrolyte;
a storage unit that stores pressure state information corresponding to the waveform; and
a pressure determination unit that determines a pressure state of the subject,
the pressure determination unit determines the pressure state of the subject by comparing the waveform acquired from the detection unit with the pressure state information read from the storage unit.
17. The pressure determination device according to claim 16, wherein the pressure determination portion includes: a counting unit that counts the number of particles contained in the body fluid that have passed through the through-hole unit due to electrophoresis, based on the waveform; and a comparison unit for comparing the number with the pressure state information,
the pressure state information includes information on a relationship between the number of the microparticles contained in the body fluid and the pressure intensity.
18. The pressure determination device according to claim 17, further comprising:
a life information input unit for inputting life information of the subject;
a life information determination unit that determines a pressure input degree based on the life information; and
a learning section that updates the stress state information based on the life information,
the learning section includes: a coincidence degree determination unit configured to determine a coincidence degree between a determination result of the pressure input degree and a determination result of the pressure state; and an updating unit that updates the relationship information based on the degree of matching, and learns the pressure state information in which the degree of matching is higher by repeating the updating by the updating unit.
Technical Field
The present invention relates to a virus detection method, a virus detection device, a virus determination program, a pressure determination method, and a pressure determination device.
Background
Patent document 1 describes the following method: the concentration of cortisol (cortisol) in saliva is measured, and the presence or absence of chronic stress in a subject is determined based on the concentration. And exemplifies the determination of cortisol concentration by liquid chromatography.
Patent document 2 describes the following method: the amount of human herpesvirus in the body fluid of the subject was measured to evaluate the degree of fatigue associated with daily life and disease. In this evaluation method, if the viral load of human herpesvirus in body fluid is large, the subject is evaluated to be in a chronic fatigue state. Examples of human herpesviruses include human herpesvirus type 6, human herpesvirus type 7, human cytomegalovirus (human cytomegalovirus) and Epstein-Barr virus (EBV). As the body fluid, blood, saliva, cerebrospinal fluid and urine can be exemplified. As a method for measuring the amount of virus, a method for measuring the amount of viral nucleic acid by a PCR method is exemplified. As described in patent documents 1 and 2, the stress and fatigue degree (hereinafter, these are collectively referred to as stress) of the subject can be evaluated by detecting the body fluid component of the subject.
Patent document 3 describes a single particle analyzer and an analysis method. The measurement container of the single particle analyzer includes a first chamber and a second chamber divided by an insulating partition wall having a through hole communicating the first chamber and the second chamber. The measurement container is provided with a first electrode which is grounded and exposed in the first chamber, and a second electrode which is grounded and exposed in the second chamber. A current meter and a power supply are interposed between the electrode exposed in the second chamber and the ground. The single particle analysis device measures the shape of the particle by measuring a detection signal between the first electrode and the second electrode when the particle contained in the liquid filled in the first chamber passes through the through hole.
Patent document 4 describes biomarkers of stress diseases detected in a sample consisting of a biological fluid of urine, blood, saliva, or cerebrospinal fluid collected from a mammal suffering from a stress disease, and one example of a stress disease is irritable bowel syndrome (irritable bowel syndrome).
Patent document 5 describes a method and a composition for treating a disease caused by herpes virus. Patent document 5 describes that herpes viruses may cause autoimmune or inflammatory diseases.
Non-patent document 1 describes a questionnaire for mood anxiety disorder in which stress is evaluated by a questionnaire (inquiry) method. The anxiety disorder questionnaire is called the so-called K6 questionnaire. The K6 questionnaire is proposed for the purpose of screening for mental diseases such as depression and anxiety (so-called mental health states), and is widely used as an index representing the degree of some mental problems including psychological stress in surveys targeted for general residents. The critical values for dividing the presence or absence and the degree of mental problems are four intervals as follows: negative is 0-4 points; the light degree is 5-8 min; the medium degree is 9-12 points; the gravity is 12-24 minutes.
Documents of the prior art
Patent document
Patent document 1: japanese patent laid-open No. 2000-275248
Patent document 2: international publication No. 2006/006634
Patent document 3: international publication No. 2013/137209
Patent document 4: japanese laid-open patent publication No. 2012-047735
Patent document 5: japanese Kohyo Table 2018-517757
Non-patent document
Non-patent document 1: kessler RC, Andrews G, COLPE LJ, et al, short screening scales to monitor publication and trees in non-specific pathological medical discovery, psychological medical 2002; 32(6): 959-76
Disclosure of Invention
Technical problem to be solved by the invention
As described in patent documents 1 and 2, stress determination methods use, for example, cortisol and herpes virus as so-called stress markers. Detection of these pressure markers requires the use of a time-consuming and labor-consuming detection method such as liquid chromatography or PCR. In addition, these detection methods have a problem of detection limit. For example, even in the PCR method which is considered to be highly sensitive, if 40 or more viruses are not present per 1. mu.l, detection is impossible, which is inconvenient. In addition, there are also problems as follows: it is impossible to detect the disease before the disease becomes a severe state (a state in which the virus is increased), and it is impossible to sufficiently perform early diagnosis and early treatment of the disease (a state of intense stress or a stress disease).
The pressure evaluation method by the questionnaire method described in non-patent document 1 has a problem that the result varies depending on the self-report and the influence of the sensitivity of the subject. Therefore, the disease sometimes cannot be appropriately diagnosed. For example, there is a problem that a stress state which is not noticed by the person is not reflected in the evaluation result. Therefore, it is desirable to provide a simple, quick, and accurate pressure determination method.
As described in patent document 4, stress has a direct relationship with a disease. It is generally known that excessive stress can lead to disease. In addition, the disease itself is generally known to cause physical and mental stress. As described in patent document 5, various diseases may be caused by herpes virus. Therefore, it is desirable to identify pathological stress conditions, stress disorders and various diseases associated with or associated with them by determining the type of virus, detecting (counting) the number of viruses, or to provide methods of identification.
The present invention has been made in view of the above circumstances, and an object thereof is to provide a virus detection method, a virus detection device for implementing the virus detection method, a virus determination program, a stress determination method, and a stress determination device for implementing the stress determination method, which can easily and quickly determine or identify stress, a stress state, a stress disease, and various diseases related to or associated with the stress, the stress state, the stress disease, and the various diseases.
Means for solving the problems
In order to achieve the above object, a virus detection method according to the present invention is characterized by comprising the steps of:
a contact step of bringing a liquid sample containing a body fluid of a subject into contact with an electrolytic solution via a through hole portion provided in a partition wall;
a current detection step of applying a voltage to the liquid sample and the electrolytic solution to the through hole portion to obtain a waveform of an ion current flowing through the through hole portion; and
a virus determination step of determining a virus type contained in the body fluid based on the waveform,
and a virus determination step of comparing the waveform with waveform information corresponding to a known virus acquired in advance to determine the type of the virus within 10 minutes.
When a virus is contained in the body fluid, according to the above configuration, when a voltage is applied to the liquid sample and the electrolyte, the waveform of the ion current flowing through the hole changes. According to the above configuration, the type of virus contained in the body fluid of the subject (patient) is determined within 10 minutes based on the change in the waveform of the ion current and waveform information corresponding to a known virus.
According to the above configuration, since the type of virus can be determined based on the waveform of the ion current corresponding to at least one type (one type) of virus, the type of virus can be determined more rapidly and accurately with high sensitivity than in the PCR method.
The concept of the virus type to be determined includes differences in "activity (status)" of viruses of the same type and model, in addition to the virus type, that is, "type" such as a difference between herpes virus and influenza virus and "model" such as a difference between herpes simplex virus type 1 and human herpes virus type 6. The concept of activity includes a so-called live-dead state of virus, for example, an infectious state (referred to as an active (viable) state or an infectious virus (infectious virus) and the like); the state of inactivation/loss of infectivity (referred to as non-active (non-viable), non-viable (non-viable), non-infectious (non-viable), etc.).
By determining the type of virus, a doctor can acquire information for identifying a disease of a subject. Further, by determining the type of virus, information for a doctor to grasp the disease state of a subject can be acquired by counting the number of viruses for each type of virus.
In addition, by determining the type of virus within 10 minutes, a doctor can be assisted in accelerating disease identification. By speeding up the disease identification, the doctor can prevent the disease of the subject from becoming serious and reduce the burden (e.g., waiting time) on the subject. In addition, the effect of treatment by doctors can be improved, and early healing can be realized.
In the above configuration, waveform information corresponding to a known virus is acquired in advance. Therefore, according to the above configuration, the type of virus contained in the body fluid of the subject can be determined by comparing the newly acquired waveform with the waveform information acquired in advance. For example, the newly acquired waveform is compared with waveform information of a known virus acquired in advance, and if they match, it is determined that there is a virus of the same type as the known virus. The newly acquired waveform is compared with the waveform information of the known virus acquired in advance, and if they do not match, it is determined that the virus is not the same virus species as the known virus. As the waveform information corresponding to the known viruses, waveform information corresponding to a plurality of known viruses may be acquired in advance.
Another feature of the virus detection method according to the present invention is that the current detection step includes: a passing step of passing the microparticles contained in the body fluid through the through-hole portion by electrophoresis.
According to the above configuration, the microparticles contained in the body fluid, that is, the microparticles contained in the liquid sample move due to electrophoresis and pass through the through-hole portion. When the fine particles pass through the through-hole portion, the waveform of the ion current corresponding to each virus changes. Based on the waveform (change in waveform) of the ion current when the fine particles pass through the through-hole portion and waveform information corresponding to known viruses, the type of the viruses contained in the body fluid of the subject is determined within 10 minutes.
According to the above configuration, by comparing the waveform obtained when the microparticles pass through the through-hole portion with the waveform information obtained in advance, the type of virus contained in the body fluid of the subject can be determined. For example, the newly acquired waveform is compared with the waveform information of the known virus acquired in advance, and if they match, it is determined that the microparticle corresponding to the newly acquired waveform is of the same virus species as the known virus. The newly acquired waveform is compared with the waveform information of the known virus acquired in advance, and if they do not match, it is determined that the microparticle corresponding to the newly acquired waveform is not of the same virus species as the known virus.
Another characteristic feature of the virus detection method according to the present invention resides in that the known virus is herpes virus.
With the above configuration, it is possible to determine whether or not the microparticle corresponding to the newly acquired waveform is a herpesvirus.
The virus detection method according to the present invention is characterized in that the body fluid is selected from the group consisting of tears and intraocular fluid.
According to the above configuration, the kind of virus leaking out into tears or intraocular fluid can be identified, and the identified kind of virus can be provided as information for diagnosing a disease caused by the virus. This information is particularly useful as information for diagnosing eye diseases, stress states and stress diseases. Tear fluid is a body fluid that can be easily and non-invasively collected. Therefore, in the case of collecting tears as a body fluid, the burden on the subject at the time of sample collection can be reduced. Here, the intraocular fluid is a fluid that fills the eyeball. The concept of ocular fluid includes at least fluid that fills the chamber of the eye and vitreous (vitreous humor).
Another feature of the virus detection method according to the present invention is that the current detection step includes: a separation step of separating viruses contained in the body fluid and foreign substances other than viruses contained in the body fluid by electroosmotic flow.
For example, when microparticles contained in a body fluid are moved by electrophoresis, if the moving distance is extended (a long flow path serving as a moving path is ensured), viruses having a large negative surface charge can be separated from microparticles having a small negative surface charge. With such a structure for separating viruses contained in body fluid and foreign substances other than viruses contained in body fluid by electroosmotic flow, among fine particles contained in body fluid, particularly viruses having a large negative surface charge selectively pass through the through-hole portion. On the other hand, other particles having a small negative surface charge hinder the passage of the through hole portion by generating an electroosmotic flow, which is a flow in the opposite direction to the particle moving direction, in the through hole portion.
Another feature of the virus detection method according to the present invention is that a disease is identified based on the type of virus determined in the virus determination step.
Based on the above structure, one or more diseases presumed from the determined virus species are identified. Through the identification of diseases, doctors can be assisted in diagnosing and treating the diseases.
In order to achieve the above object, a virus detection device according to the present invention includes: a sample storage unit for storing a liquid sample containing a body fluid of a subject; an electrolyte storage unit that stores an electrolyte; a partition wall portion that partitions the sample storage portion and the electrolyte solution storage portion; a through hole portion provided in the partition wall portion and communicating the sample storage portion and the electrolyte solution storage portion; a detection unit that obtains a waveform of an ion current flowing through the through hole unit by applying a voltage to the liquid sample and the electrolyte; a storage unit that stores a learned model that has been machine-learned so that, when the waveform is input, a virus type corresponding to waveform information of a known virus type is output; and a virus determination unit that applies the waveform acquired from the detection unit to the learned model read from the storage unit to determine a virus type contained in the body fluid.
According to the above configuration, the above virus detection method can be realized. That is, when a voltage is applied to the liquid sample stored in the sample storage portion and the electrolyte stored in the electrolyte storage portion, an ion current flows through the through hole portion of the partition wall portion. By detecting the ion current by the detection unit, the waveform of the ion current corresponding to the fine particles (viruses) contained in the body fluid of the subject can be obtained. When the waveform is acquired, the virus determination unit applies the waveform acquired by the detection unit to the learned model to accurately determine the type of virus contained in the body fluid. Note that the learned model is an example of waveform information corresponding to a virus. In addition, applying a waveform to a learned model is an example of a comparison of a waveform with waveform information.
By determining the type of virus, a doctor can acquire information for identifying a disease of a subject. Further, by determining the type of virus, information for a doctor to grasp the disease state of a subject can be acquired by counting the number of viruses for each type of virus.
According to the above configuration, the accuracy of the outputted virus type can be improved by enhancing machine learning of the learned model. Therefore, the determination speed of virus determination can be easily increased. For example, it is possible to realize a high speed of judging the virus type within 10 minutes. This can assist the doctor in expediting the disease identification. By accelerating the identification of a disease, the severity of the disease can be prevented, the effect of a doctor on the treatment can be improved, and early healing can be achieved.
In order to achieve the above object, a virus detection device according to the present invention includes: a sample storage section that stores a liquid sample containing a body fluid of a subject selected from the group consisting of tears and intraocular fluid;
an electrolyte storage unit that stores an electrolyte; a partition wall portion that partitions the sample storage portion and the electrolyte solution storage portion; a through hole portion provided in the partition wall portion and having hydrophilicity for communicating the sample storage portion and the electrolyte storage portion; a detection unit that obtains a waveform of an ion current flowing through the through hole unit by applying a voltage to the liquid sample and the electrolyte; a storage unit for storing waveform information of a known virus type; and a virus determination unit that determines a virus type contained in the body fluid based on the waveform, wherein the virus determination unit determines the virus type contained in the body fluid by comparing the waveform acquired from the detection unit with the waveform information of the known virus type read from the storage unit.
According to the above configuration, the above virus detection method can be realized in the case of using a liquid sample containing a body fluid of a subject selected from a tear fluid or an intraocular fluid. That is, since the through-hole portion has hydrophilicity, the through-hole portion can be quickly filled with the liquid sample, and the conduction current can be conducted. When a voltage is applied to the liquid sample stored in the sample storage portion and the electrolyte stored in the electrolyte storage portion, an ion current flows through the through hole portion of the partition wall portion. By detecting the ion current by the detection unit, the waveform of the ion current corresponding to the fine particles (viruses) contained in the body fluid of the subject can be obtained. By comparing the waveform with the waveform information of known viruses stored in the storage unit, the type of virus contained in the body fluid of the subject can be accurately determined.
According to the above configuration, the waveform of the ion current changes for each virus (each virus) when passing through the through hole portion, and therefore, the type of virus can be determined at a higher speed and with higher sensitivity than in the PCR method.
The virus determination unit compares, for example, a newly acquired waveform with waveform information of a known virus acquired in advance, and if they match, determines that the microparticle corresponding to the newly acquired waveform is of the same virus type as the known virus. The virus determination unit compares the newly acquired waveform with waveform information of a known virus acquired in advance, and if they do not match, determines that the particle corresponding to the newly acquired waveform is not of the same virus type (virus type) as the known virus. As the waveform information corresponding to the known viruses, the waveform information corresponding to a plurality of known viruses may be stored in the storage unit.
Another feature of the virus detection device according to the present invention is that the liquid sample contains glutathione (Ox iglutatine).
According to the above configuration, the activity of the virus contained in the body fluid of the subject can be enhanced or the reduction of the virus activity can be prevented. As a result, the accuracy of determining the type of virus contained in the body fluid of the subject is improved.
Another characteristic configuration of the virus detection apparatus according to the present invention is that the virus determination unit determines the type of the virus within 10 minutes, and is portable.
According to the above configuration, the virus detection device is portable. Thus, the doctor can freely carry the virus detection device to a desired place when necessary. Therefore, when a patient as a subject appears, the type of virus can be determined at a place where the examination burden on the patient is small, and a doctor can be assisted in diagnosis and treatment of a disease.
According to the above configuration, the type of virus can be determined within 10 minutes, thereby assisting a doctor in accelerating disease identification. By accelerating the identification of a disease, the severity of the disease can be prevented, the effect of a doctor on the treatment can be improved, and early healing can be achieved.
In order to achieve the above object, a virus determination program according to the present invention for causing a computer to execute a process of determining a virus type contained in a body fluid of a subject includes: a reception process of receiving a waveform of an ion current obtained by applying a voltage to a liquid sample containing the body fluid and an electrolytic solution; a virus determination process of determining a virus type contained in the body fluid based on the waveform received in the reception process; and a transmission process of transmitting a determination result in the virus determination process, the virus determination process determining a type of a virus based on a learned model subjected to machine learning so that, when the waveform is input, a virus type corresponding to waveform information of a known virus type is output.
According to the above configuration, the virus detection device, which is a computer (hereinafter, simply referred to as a CPU), can be caused to execute a process of determining the type of virus contained in the body fluid of the subject. That is, according to the above configuration, the CPU can execute the reception process of the waveform of the ion current, the virus determination process of determining the type of the virus contained in the body fluid based on the received waveform, and the transmission process of transmitting the determination result.
According to the above configuration, when the CPU is caused to execute the virus determination process, the CPU is caused to execute a process of applying the received waveform to the learned model to determine the type of the virus contained in the body fluid. This makes it possible to determine the type of virus based on the learning result of machine learning.
In order to achieve the above object, a pressure determination method according to the present invention includes the following steps: a contact step of bringing a liquid sample containing a body fluid of a subject into contact with an electrolytic solution via a through hole portion provided in a partition wall; a current detection step of applying a voltage to the liquid sample and the electrolytic solution to the through hole portion to obtain a waveform of an ion current flowing through the through hole portion; and a determination step of comparing the waveform with specific pressure state information to determine the pressure state of the subject.
Various substances leak into the body fluid of the subject depending on the pressure state of the subject. Hereinafter, a substance that leaks out according to the pressure state of the subject is referred to as a pressure mark. The term "body fluid" refers to a liquid that naturally oozes out of or is discharged from the human body, such as tears and saliva. According to the above configuration, the leakage state of the pressure mark can be obtained as a waveform of the ion current (hereinafter, may be simply referred to as a waveform). That is, by applying a specific voltage to the liquid sample and the electrolyte, a specific pressure mark is moved from the liquid sample to the electrolyte. When the pressure mark passes through the through-hole portion, an ion current flowing between the liquid sample and the electrolytic solution changes. Thereby, a waveform corresponding to the leaking state of the pressure mark can be acquired. The waveform thus obtained corresponds to the magnitude of the pressure of the subject. Acquisition of the waveform can be performed extremely easily and quickly as compared with an analysis method such as a PCR method or a liquid chromatography method.
The pressure state information is information on a relationship between a waveform of a specific ion current and a pressure state (for example, a state of pressure accumulation) of a human. The stress state is an index, such as a stress score, objectively grasped from a work environment (for example, a human occupation, a job title, a duty, a work time period, and the like). Therefore, according to the above configuration, the pressure state of the subject can be determined by comparing the waveform with the specific pressure state information. The determination is accurate because it is not affected by the sensitivity as reported by the self.
Another characteristic structure of the pressure determination device according to the present invention is characterized in that the current detection step includes: a passage step of electrophoretically passing the microparticles contained in the body fluid through the through-hole portion, the pressure determination step including: a counting step of counting the number of the fine particles passing through the through hole portion based on the waveform; and a comparison step of comparing the number of the particles with the pressure state information; the pressure state information includes information on a relationship between the number of the microparticles contained in the body fluid and the pressure intensity.
According to the above configuration, microparticles (for example, herpes virus) contained in the body fluid can be moved by electrophoresis and pass through the through-hole portion. Since the waveform does not change during the passage, the number of microparticles contained in the body fluid of the subject can be counted based on the change in the waveform.
Further, according to the above configuration, if the information on the relationship between the pressure intensity and the number of microparticles contained in the body fluid of the person (for example, the calibration curve) is acquired in advance as the pressure state information, the pressure intensity of the subject can be determined based on the number of microparticles contained in the body fluid of the subject.
Another characteristic structure of the pressure determination method according to the present invention is that the microparticles are herpesviruses.
Many adults suffer from herpes viruses. When the pressure of a human increases, the herpes virus leaks out into the body fluid due to a change in the state of the immune system accompanying the increase in pressure. Therefore, according to the above configuration, the number of herpes viruses in the body fluid of the subject is directly detected, whereby the stress state of the subject can be easily and quickly determined. In addition, since the device is not influenced by the sensitivity as in self-declaration, the device is accurate.
Another characteristic structure of the pressure determination method according to the present invention is that the body fluid is tear fluid.
Tears are body fluids that can be easily and non-invasively collected. Therefore, the burden on the subject can be reduced. For example, a pipette or the like may be used to directly collect tears, or a cleaning solution containing tears for cleaning the eye may be collected.
In addition, since the fine particles in the tear fluid favorably correspond to the pressure state of the subject, the detection sensitivity is improved. For example, herpes viruses latently infect ganglia of the body, such as the trigeminal ganglia. The trigeminal nerve is a kind of cranial nerve, and an ophthalmic nerve that innervates a human eye is included in the trigeminal nerve. Thus, herpes viruses that reside in the trigeminal ganglion are prone to leakage into tears that leak from the lacrimal gland of the eye. Further, when the pressure intensity of the subject becomes high, more herpes virus leaks into the tear fluid. Therefore, according to the above configuration, by using tear fluid as the body fluid, the pressure can be determined easily and quickly.
In order to achieve the above object, a pressure determination device according to the present invention includes: a sample storage unit for storing a liquid sample containing a body fluid of a subject; an electrolyte storage unit that stores an electrolyte; a partition wall portion that partitions the sample storage portion and the electrolyte solution storage portion; a through hole portion provided in the partition wall portion and communicating the sample storage portion and the electrolyte solution storage portion; a detection unit that obtains a waveform of an ion current flowing through the through hole unit by applying a voltage to the liquid sample and the electrolyte; a storage unit that stores pressure state information corresponding to the waveform; and a pressure determination unit that determines a pressure state of the subject, wherein the pressure determination unit determines the pressure state of the subject by comparing the waveform acquired from the detection unit with the pressure state information read from the storage unit.
According to the above configuration, the pressure determination method can be realized. That is, when a voltage is applied to the liquid sample stored in the sample storage portion and the electrolyte stored in the electrolyte storage portion, an ion current flows through the through hole portion of the partition wall portion. By detecting the ion current by the detection unit, the leakage state of the pressure mark can be obtained in the form of the waveform of the ion current. By comparing the waveform with the pressure state information, the pressure state of the subject can be determined.
Another characteristic structure of the pressure determination device according to the present invention is characterized in that the pressure determination unit includes: a counting unit that counts the number of particles contained in the body fluid that have passed through the through-hole unit due to electrophoresis, based on the waveform; and a comparing unit that compares the number with the pressure state information, the pressure state information including information on a relationship between the number of the microparticles contained in the body fluid and the pressure intensity.
When a voltage is applied to the liquid sample stored in the sample storage portion and the electrolyte stored in the electrolyte storage portion, particles contained in the body fluid move due to electrophoresis. When the particles moved by the electrophoresis pass through the through-hole portion, the waveform of the ion current changes. The counting unit counts the number of particles passing through the through hole portion based on the change in the waveform of the ion current. The comparison unit compares the number counted by the counting unit with the pressure state information. Here, since the pressure state information includes information on the relationship between the number of microparticles contained in the body fluid and the pressure intensity, the pressure determination unit can determine the pressure state of the subject by comparing the pressure state information with the pressure intensity of the body fluid.
Another characteristic structure of the pressure determination device according to the present invention is characterized by further comprising: a life information input unit for inputting life information of the subject; a life information determination unit that determines a pressure input degree based on the life information; and a learning unit that updates the stress state information based on the life information, the learning unit including: a coincidence degree determination unit configured to determine a coincidence degree between a determination result of the pressure input degree and a determination result of the pressure state; and an updating unit that updates the relationship information based on the degree of coincidence, and learns the pressure state information in which the degree of coincidence is higher by repeating the updating by the updating unit.
According to the above configuration, the coincidence degree determination unit of the learning unit determines the coincidence degree between the determination result of the pressure input degree based on the life information such as the working environment and the determination result of the pressure state based on the ion current waveform. The updating unit of the learning unit updates the relationship information based on the matching degree. The learning unit learns the pressure state information in which the degree of coincidence between the result of the determination of the pressure state and the result of the determination of the degree of pressure input is high by repeating the update of the relationship information by the updating unit. Thus, a pressure determination device in which the determination accuracy of the pressure determination unit based on the waveform and the pressure state information is improved can be provided.
ADVANTAGEOUS EFFECTS OF INVENTION
The invention can provide a simple, rapid and accurate virus detection method, virus detection device, virus determination program, pressure determination method and pressure determination device.
Drawings
Fig. 1 is a schematic configuration diagram of a determination device according to a first embodiment.
Fig. 2 is a waveform diagram in the case where no peak exists.
Fig. 3 is a waveform diagram in the presence of a peak.
Fig. 4 is a waveform diagram showing an example of a peak.
Fig. 5 is an explanatory view of the shape of a peak.
FIG. 6 is an explanatory view of the structure of the partition wall and the through hole and the virus electrophoresis.
Fig. 7 is a diagram showing a relationship among jobs, passage numbers, and stress evaluations.
Fig. 8 is a diagram showing the style of the K6 questionnaire.
Fig. 9 is a schematic configuration diagram of a determination device according to the second embodiment.
Fig. 10 is a graph showing a relationship between the degree of pressure input and the number of passing particles.
Fig. 11 is a diagram showing an example of the shape of the peak of each virus species.
Fig. 12 is a schematic configuration diagram of a detection device according to a third embodiment.
Fig. 13 is an illustration of the shape of a peak.
Fig. 14 is an illustration of the shape of a peak.
Fig. 15 is an illustration of the shape of a peak.
Figure 16 is an example of a waveform obtained from a sample of a patient with HSV corneal herpes.
FIG. 17 is an example of a waveform obtained from a sample of a patient with CMV intracorneal dermatitis.
Fig. 18 is a schematic configuration diagram of a detection device according to a fourth embodiment.
Fig. 19 is a flow of an operation performed by the determination device according to the virus determination program.
Detailed Description
A pressure determination method, a pressure determination device and a virus detection method for realizing the pressure determination method, a virus detection device, and a virus determination program according to embodiments of the present invention will be described below with reference to the drawings.
[ first embodiment ]
[ about samples ]
In the present embodiment, a case where tear liquid is used as the body fluid of the subject will be described by way of example. As the tears, a cleansing solution containing tears collected by cleansing the conjunctival sac (bag-like portion at the boundary between the eyeball and the lower eyelid) of a subject with a phosphate buffered saline (hereinafter, referred to as a physiological saline) is used. Hereinafter, this cleaning liquid is referred to as a sample L1. Namely, sample L1 is a physiological saline solution containing tears. The sample L1 is an electrolyte solution, and an ion current can be conducted by common salt (sodium chloride). It is stated that there is often 7 microliters of tear fluid in the conjunctival sac. In the present embodiment, the conjunctival sac is washed with 30 μ L of physiological saline.
It is known that herpes viruses leak into tears of a subject in accordance with the pressure state of the subject. Herpes viruses are fine particles having a particle size (diameter) of approximately 200 nm. For example, the particle size of herpes simplex virus type 1 (HSV-1 (also abbreviated as HHV1)) is approximately 150-180 nm. The particle size of varicella/zoster virus (VZV (also abbreviated as HHV3)) is approximately 180-200 nm. The particle size of human cytomegalovirus (HCMV (also abbreviated as HHV5)) is approximately 150-200 nm. The particle size of human herpes virus type 6 (HHV-6) is approximately 200 nm.
Herpes viruses have an envelope formed of a lipid bilayer membrane on the outside of a nucleocapsid (nucleocapsid) composed of a viral nucleic acid in the center of a microparticle and a protein capsid surrounding the viral nucleic acid. The glycoprotein called spike protein (spike) protrudes to the surface of the herpes virus envelope. Amino acids having negatively charged side chains are contained in the glycoprotein. Therefore, the surface potential of the herpesvirus particles is negative in body fluid and physiological saline.
Hereinafter, the present embodiment will be described by exemplifying a case where at least herpes simplex virus type 1 is included as a herpes virus. In addition, the following description will also exemplify a case where herpes virus leaking into tear fluid is used as a pressure marker of a subject. Hereinafter, herpes viruses are sometimes simply referred to as viruses.
[ concerning the pressure judging apparatus ]
Fig. 1 shows a configuration of a determination device 100 that realizes the pressure determination method according to the present invention and is used as a pressure determination device. The determination device 100 includes: an inspection substrate 1 which is an inspection chip of a liquid sample L1 (an example of a liquid sample) containing a body fluid of a subject; a detection unit 7 that acquires information on the waveform of the ion current from the inspection substrate 1; a control unit 10, such as a computer, for determining the pressure state of the subject based on the information acquired from the inspection substrate 1; and a power supply section 71. The detection unit 7 and the like are connected to the control unit 10 via a network N such as a data bus or a computer network (local area network or the internet) capable of bidirectional communication.
The inspection substrate 1 is a plate-like substrate made of an insulator such as quartz glass or polydimethylsiloxane. The inspection substrate 1 includes: a sample reservoir 2 that stores a sample L1, an electrolyte reservoir 3 that stores an electrolyte L2, a partition wall 5 (an example of a partition wall portion) in which a through hole 6 (an example of a through hole portion) is formed, a first electrode 72, and a second electrode 73 that is paired with the first electrode 72. The inspection substrate 1 was used as a detection chip of the sample L1. As the electrolyte solution L2, an electrolyte solution such as a physiological saline solution is used. The electrolyte L2 can conduct an ion current through common salt (sodium chloride).
The sample storage section 2 is a part serving as a container for storing the sample L1. For example, a concave portion is provided on one surface of the inspection substrate 1. The sample storage section 2 includes: a first circular recess 20 which is a circular recess and is the main storage container of the sample L1; and a first flow channel 21 which communicates with the through hole 6 of the partition wall 5 from the first circular recess 20. The first flow path 21 is connected to one surface of the partition 5. A first electrode 72 is provided between the first circular recess 20 and the partition 5 in the first channel 21. The sample storage section 2 is filled with a sample L1. When the sample reservoir 2 is filled with the sample L1 without bubbles, the first electrode 72 and the sample L1 are electrically connected. The liquid-receiving surface of the sample storage section 2 is preferably surface-treated to make it hydrophilic. As the surface treatment, for example, plasma treatment, ultraviolet irradiation, chemical modification, or the like can be used.
The electrolyte solution storage unit 3 is a part serving as a container for storing the electrolyte solution L2. For example, a concave portion is provided on the other surface of the inspection substrate 1. The electrolyte reservoir 3 includes: a second circular recess 30 which is a circular recess and is a main storage container of the electrolyte L2; and a second channel 31 that communicates with the through hole 6 of the partition wall 5 from the second circular recess 30. The second channel 31 is connected to the other surface of the partition 5 to which the first channel 21 is connected. A second electrode 73 is provided between the second circular recess 30 and the partition wall 5 in the second channel 31. Electrolyte L2 is filled in electrolyte reservoir 3. When the electrolyte L2 is filled in the electrolyte reservoir 3 without bubbles, the second electrode 73 is electrically connected to the electrolyte L2. Further, the sample L1 and the electrolyte L2 were physically connected via the through-hole 6. When the sample L1 and the electrolyte L2 are physically connected, conduction can be established between the sample L1 and the electrolyte L2.
The first electrode 72 is electrically connected to one end of a power supply section 71 serving as a dc power supply. A specific dc voltage (for example, 0.1 v) is applied from the power supply section 71 to the first electrode 72. In the present embodiment, the first electrode 72 is a negative electrode. As the first electrode 72, for example, a silver/silver chloride electrode in which the surface of silver is covered with silver chloride can be used. Hereinafter, the voltage applied from the power supply section 71 to the first electrode 72 may be simply referred to as an applied voltage.
The other end of the power supply section 71 is connected to the ground G. The power supply unit 71 is communicably connected to the control unit 10 via the network N. The power supply section 71 applies a specific voltage in accordance with an instruction from the control section 10.
The second electrode 73 is an electrode having a polarity opposite to that of the first electrode 72. In the present embodiment, the second electrode 73 is a positive electrode. The second electrode 73 is electrically connected to the ammeter 70 of the detection unit 7. A potential difference of direct current is generated between the first electrode 72 and the second electrode 73 by the voltage applied from the first electrode 72. As the second electrode 73, a silver/silver chloride electrode can be used as in the case of the first electrode 72. Hereinafter, the dc potential difference generated between the first electrode 72 and the second electrode 73 is simply referred to as a potential difference. The viruses of the sample L1 move toward the second electrode 73 along the electric field formed by the potential difference (electrophoresis). An ion current flows between the first electrode 72 and the second electrode 73 due to the voltage applied from the first electrode 72. The ion current flows through the through-hole 6.
The detection unit 7 is a detection unit having an ammeter 70 and an interface (not shown) for transmitting waveform information of the current value acquired by the ammeter 70 to the control unit 10 via the network N. The ammeter 70 is a detector that detects a current flowing between the second electrode 73 and the ground G. The ion current flowing between the first electrode 72 and the second electrode 73 is discharged from the second electrode 73 to the ground G via the ammeter 70. The detection unit 7 acquires waveform information of the ion current based on the current value information of the ion current acquired by the ammeter 70, and transmits the waveform information to the control unit 10. Hereinafter, the waveform information of the ion current may be simply referred to as a waveform, and particularly, a waveform obtained by storing the sample L1 in the sample storage unit 2 may be referred to as a sample waveform. Fig. 2 and 3 show examples of waveforms of changes in the current value of the ion current with the lapse of time t (sec) when a specific voltage is applied from the first electrode 72.
As shown in fig. 6, the partition wall 5 includes: made of silicon nitride (Si)3N4) A substrate 50 in the form of a thin plate (thin film) made of an inorganic oxide solid such as an insulator, a reinforcing plate 51 made of a solid of a semiconductor such as silicon (Si) to which the substrate 50 is attached, and a through-hole 6 formed in the substrate 50. The substrate 50 is disposed on the side in contact with the first flow path 21. The thickness of the substrate 50 is, for example, 50 nm. The thickness of the reinforcing plate 51 is, for example, 500 μm. The partition wall 5 is obtained by coating the surface of a reinforcing plate 51 with silicon nitride to form a substrate 50.
The through-hole 6 is a fine pore (solid hole) penetrating the solid substrate 50. In the present embodiment, the diameter of the through-hole 6 is larger than the diameter of the virus, and is, for example, a circular hole (so-called nanopore) having a diameter of 300 nm. The diameter of the through hole 6 is formed larger than the thickness of the substrate 50. The through-hole 6 is a low aspect ratio nanopore having an aspect ratio smaller than 1 in terms of the aspect ratio (the ratio of the depth of the substrate 50 to the diameter of the hole).
The inner surface 6a of the through-hole 6 is subjected to surface treatment so as to have a negative surface charge of the same polarity as that of the virus. Further, the inner surface 6a is also subjected to surface treatment to make it hydrophilic. For example, the inner surface 6a may be formed of a surface treatment agent that is molecularly modified with sugar chains present on the surface of the virus to be detected. Since the inner surface 6a has negative surface charges, sodium ions (cations) of the sample L1 and the electrolytic solution L2 are attracted to the vicinity of the surface of the inner surface 6a, forming an electric double layer. As the surface treatment for imparting hydrophilicity to the inner surface 6a, for example, chemical modification such as molecular modification, plasma treatment, ultraviolet irradiation treatment, and the like can be used.
Note that, a window hole 61, which does not block the opening portion of the through-hole 6 and has a diameter larger than that of the through-hole 6, is formed in a portion of the reinforcing plate 51 corresponding to the through-hole 6. The through-hole 6 penetrates the partition wall 5 through the window hole 61.
A method of forming the through-hole 6 will be explained. The through-hole 6 may be formed by: a part of the reinforcing plate 51 is subjected to anisotropic etching of silicon using an aqueous solution such as potassium hydroxide to open the window hole 61, a part of the substrate 50 corresponding to the window hole 61 is made into a silicon nitride film, and then a micropore is drawn in the part by an electron beam drawing method or the like, and the drawn part is excavated by a reactive ion etching method or the like.
The waveform of the ion current and the electrophoresis of the virus are generally described. As shown in fig. 6, when a voltage is applied to the first electrode 72, the virus V of the sample L1 moves toward the second electrode 73 by electrophoresis due to the potential difference. Due to this movement, the virus V passes through the through-hole 6.
Further, an electroosmotic flow EF is generated in the vicinity of the inner side of the electric double layer of the inner surface 6a due to a potential difference caused by the voltage application to the first electrode 72. Since the inner surface 6a has a negative surface charge of the same polarity as the virus, the flow direction of the electroosmotic flow EF is opposite to the moving direction of the virus V. Due to the electroosmotic flow EF, the foreign particles P (for example, particles such as proteins) having a negative surface charge smaller than that of the virus V inhibit the movement of the virus V in the moving direction. Therefore, the inclusion particles P cannot pass through the through-holes 6. Thereby, the inclusion particles P and the virus V are separated, and the virus V selectively passes through the through-holes 6 (an example of the separation step).
When a voltage is applied to the first electrode 72, an ion current flows between the first electrode 72 and the second electrode 73. The current value of the ion current depends on the concentrations of the electrolyte (sodium chloride) in the sample L1 and the electrolyte solution L2, and the cross-sectional area of the cross section of the electrolyte solution (the sample L1, the electrolyte solution L2, and a mixture thereof) existing in the hole of the through-hole 6, the cross-sectional area intersecting the axial direction of the hole of the through-hole 6. Therefore, when particles such as viruses enter the through-hole 6, the electrolyte in the hole of the through-hole 6 is repelled, and thus the ion current flowing through the through-hole 6 is reduced. That is, one spike-like waveform, i.e., a peak Pk, in which the current value decreases is generated in the waveform of the ion current flowing between the first electrode 72 and the second electrode 73 at the time of passage of each virus through the through hole 6 (see fig. 3). Therefore, if the number of peaks Pk detected per unit time is counted, the number of viruses passing through the through-hole 6 per unit time can be measured (hereinafter, the number of viruses already passing through the through-hole 6 may be simply referred to as the number of passes). In the case of fig. 3, the number of peaks Pk detected after 200 seconds detection is 18, and the number of passing peaks Pk measured is 18. In the case where no fine particles such as viruses are present in the sample L1, the peak Pk is not generated in the waveform of the ion current (see fig. 2). The determination device 100 can detect viruses sharply on a 1-by-1 basis.
The peaks Pk when the virus particles of herpes simplex virus type 1 (HSV-1) pass through the through-holes 6 are illustrated in fig. 4. The wave peak Pk of fig. 4 is illustrated in an enlarged manner in fig. 5. As shown in fig. 5, the peak Pk has a specific pulse height Ip and a specific pulse width td. The pulse height Ip is a difference between a peak start point or a peak end point and a peak top in the peak Pk. The pulse width td is an elapsed time from a peak start point to a peak end point in the peaks Pk. The shape of the peak Pk such as the pulse height Ip and the pulse width td differs depending on the type of virus (virus type, virus state (activity), that is, the particle size of the virus, the type of glycoprotein on the envelope surface, and the like).
Here, the concept of the type of virus according to the present embodiment includes the difference in "activity (status)" of the same type and model of virus, in addition to the type of virus, that is, "type" such as the difference between herpes virus and influenza virus and "model" such as the difference between herpes simplex virus type 1 and human herpes virus type 6. The concept of activity includes a so-called live-dead state of virus, for example, an active/infectious state (referred to as active (viable), infectious virus (infectious) or the like); the state of inactivation/loss of infectivity (referred to as non-active (non-viable), non-viable (non-viable), non-infectious (non-viable), etc.). Hereinafter, the virus in an active/infectious state is simply referred to as "active virus" or the like. In addition, a virus in an inactivated/lost infectious state is simply referred to as "inactive virus" or the like. Active viruses and inactive viruses have different surface potentials due to, for example, a difference in the state of the particle surface (for example, herpes viruses lose their activity when the envelope is disrupted). Thus, active and inactive viruses produce distinct peaks Pk.
The control unit 10 is a functional unit that controls the entire determination device 100. The control unit 10 of the present embodiment is realized by software by a computer (in the present embodiment, a personal computer, hereinafter, simply referred to as a CPU) including at least a central processing unit and a temporary memory. The control unit 10 receives information input from an input unit 91 (an example of a life information input unit), and outputs various kinds of information processed internally from an output unit 92, the input unit 91 being an input interface for inputting information such as a keyboard and a mouse connected to the CPU; the output unit 92 is an output interface for outputting information, such as a monitor, a speaker, or a printer. Various information such as software (program) for realizing the control unit 10, information input from the input unit 91, information to be output from the output unit 92, and pressure state information for internal processing by the control unit 10 is stored in the storage unit 8 as a storage device or a storage medium. The input unit 91, the output unit 92, and the storage unit 8 can be connected to each other by communication via a network N. Examples of the storage unit 8 include, but are not limited to, magnetic storage media such as hard disks, optical disks such as CDs and DVDs, flash memories such as SSDs, USB memories, and memory cards, cloud servers, rental servers, and the like.
As shown in fig. 1, the control unit 10 includes a determination unit 11 (an example of a pressure determination unit) implemented by software. The determination unit 11 is a functional unit that determines the pressure state of the subject by comparing the waveform acquired from the detection unit 7 with the pressure state information read from the storage unit 8. The judgment unit 11 includes: a counting unit 12 that counts the number of viruses (passage number) that have passed through the through-hole 6 due to electrophoresis, the viruses being contained in the sample L1 per unit time, based on the waveform; and a comparison unit 13 for comparing the passage number with the pressure state information.
The pressure state information includes: relationship information (hereinafter, simply referred to as relationship information) between a waveform of a sample acquired under a specific condition and a pressure state (e.g., the kind and intensity of pressure) of a person (e.g., a subject). In the present embodiment, the pressure state information includes at least information on the relationship between the number of peaks Pk of the sample waveform and the pressure intensity, which are acquired in advance under a specific detection condition. The specific measurement conditions include, for example, the value of the applied voltage, the measurement time (the time during which the voltage is applied and continuously counted), the electrolyte concentrations of the sample L1 and the electrolyte solution L2, and the structure of the test substrate 1. Hereinafter, the description will be made on the premise that specific measurement conditions are satisfied.
An example of relationship information is shown in table 1. The relationship information includes: a range of the number of passes per unit time of 5 minutes, pressure intensity levels (S1 to S5) corresponding thereto, and post information corresponding to the respective pressure intensity levels. The job is exemplified by a nurse (after-day shift), a nurse (after-night shift), a ward nurse manager (ward clerk), and a ward pharmacist assigned to a hospital ward. Specifically, 5 levels of a nurse (after-shift) (weak stress), a ward nurse (after-shift) (medium stress), a ward administrator (strong stress), and a ward pharmacist (strong stress) may be respectively corresponded in the order from the level S1 to the level S5. Note that the pressure intensity levels are sequentially increased in order from the level S1 to the level S5. In addition, the duties (particularly, the cause of mental stress) and the working hours (particularly, the cause of physical stress) required for these jobs are increased in order from a nurse (after-shift), a ward nurse manager, a ward administrator, and a ward pharmacist. Note that such relationship information may be constructed by acquiring the number of passage of a plurality of subjects and the pressure intensity in advance.
TABLE 1
With respect to the relationship information of table 1, a modified example will be explained. When the number of passes is 17, the pressure intensity level corresponds to the level S4. And the job corresponds to a ward administrator. When the number of passes is 11, the pressure intensity level corresponds to the level S3. And the job corresponds to the ward nurse manager.
The counting unit 12 detects the number of peaks Pk in the waveform acquired in a specific time period (for example, a short time of about 5 minutes) in a state where the voltage is applied to the first electrode 72, and counts the number of passes by regarding the number of peaks Pk as the number of passes. Note that, if the pulse wave height Ip of the peak Pk is smaller than a certain value (for example, smaller than 0.05 nanoampere), the counting section 12 excludes the peak from the detection target.
The comparing unit 13 compares the passage number counted by the counting unit 12 with the pressure state information read from the storage unit 8 to determine the pressure intensity level corresponding to the passage number. For example, when the number of passes counted by the counting unit 12 is 17, the comparison unit 13 determines that the pressure intensity level is S4 level. When the number of passages is 11, the comparison unit 13 determines that the pressure intensity level is the S3 level. In this way, the determination unit 11 determines the pressure of the subject.
[ series of procedures for pressure determination ]
A series of flows of pressure determination will be described. The measurer or the like stores the sample L1 in the sample storage section 2. Further, the electrolyte solution L2 was stored in the electrolyte solution storage section 3. Then, the specimen L1 and the electrolytic solution L2 were brought into contact via the through-hole 6 to become an energizable state (contact step).
When a measurer or the like inputs measurement conditions (for example, detection time and applied voltage) and an instruction to start measurement to the control unit 10 via the input unit 91 after the sample L1 and the electrolyte L2 become an electrically conductive state, the control unit 10 gives an instruction to the power supply unit 71 to apply voltage under the conditions corresponding to the measurement conditions. In response to this instruction, the power supply section 71 applies a voltage to the first electrode 72 under a specific condition.
When a specific voltage is applied to the first electrode 72, the viruses of the sample L1 move toward the second electrode 73 by electrophoresis. When the virus passes through the through hole 6 during the movement, a peak Pk is generated in the waveform of the ion current (current detection step, passage step). The counting unit 12 counts the number of the peaks Pk as a passage number (determination step, counting step).
The comparing unit 13 determines the corresponding pressure intensity based on the passage number and the relationship information (pressure determining step, comparing step). The control unit 10 acquires the determination result from the determination unit 11 and outputs it to the output unit 92 (for example, displays it on a monitor). In this output, the control unit 10 intuitively reads the term that is replaced with the term that enables the pressure state to be grasped, based on the determination result and the relationship information acquired from the determination unit 11, and outputs the term from the output unit 92. For example, if the determination result is level S1, the output unit 92 displays "weak pressure". By this replacement, the user can intuitively grasp the pressure level. For example, when the determination result is level S1, the output unit 92 displays "weak pressure". By this reading, the user can intuitively grasp the pressure level.
[ example 1 ]
The number of passes (vertical axis) of the test performed on the sample L1 of the subject for each of the duties in the hospital ward is shown in fig. 7. The values obtained by 5-minute detection are shown by the numbers. The jobs exemplified 5 kinds of nurses (after shift), ward nurses, ward managers, and ward pharmacists. In fig. 7, each job is also represented by an icon of a black solid circle mark, a white open circle mark, a black solid square mark, a white open square mark, and a black solid triangle mark in the order from the nurse (after the shift) to the ward pharmacist. The pressure determination method and the pressure determination device 100 according to the present embodiment will be described below with respect to the pressure determination of the subject corresponding to these icons.
The determination unit 11 determines the pressure intensity of the subject corresponding to each icon based on the number of passes and the relationship information in table 1. In the case of fig. 7, the stress intensity levels of the subjects were determined as S1, S2, S3, S4 and S5 in the order of nurse (after shift), ward nurse manager (ward clerk) and ward pharmacist.
When these determination results are acquired from the determination unit 11, the control unit 10 replaces the reading of the determination results at the S1 level or the like with terms that allow intuitive understanding of the pressure state, and outputs the result to the output unit 92. In the example of fig. 7, the judgment results of the respective icons of the nurse (after shift), the ward nurse manager, the ward administrator, and the ward pharmacist are sequentially read and replaced with "weak pressure", "medium pressure", "strong pressure", and are output from the output unit 92 and the like.
In the case of the present embodiment, the determination result coincides with the job title of the subject. That is, the judgment result obtained by the stress judgment method according to the present embodiment matches the job and the amount of working hours required for the job of the subject. As described above, according to the stress determination method of the present embodiment, it is possible to perform evaluation with high correlation between the job and the operating time required for the job of the subject.
[ comparative example ]
Fig. 7 shows the score (horizontal axis) of stress evaluation by questionnaire for each subject of each job in the hospital ward in addition to the number of passes. Stress evaluation by the questionnaire method uses the K6 questionnaire (see fig. 8) corresponding to the anxiety disorder questionnaire described in non-patent document 1. The score of the stress evaluation (hereinafter, simply referred to as score) can be understood as: the greater this value is declared by itself, the greater the pressure experienced.
Based on the K6 questionnaire, the greater the score of the stress assessment, the greater the stress experienced. However, in the example of fig. 7, no compelling correlation is obtained between the duties and working hours required for the duties of the examinees and the scores of stress evaluations. For example, when a nurse (after work) is focused on, scores of stress evaluations for individual subjects vary greatly even with the same responsibilities and working hours.
As described above, according to the determination device 100 of the present embodiment, it is possible to easily and quickly perform evaluation with high correlation between the type and size of external factors such as the responsibility required for the job of the subject and the stress such as the work hours, as compared with the conventional questionnaire survey method.
[ second embodiment ]
The control unit 10 of the determination device 100 according to the first embodiment includes the determination unit 11. As shown in fig. 9, the determination device 100 according to the second embodiment is different in that the control unit 10 further includes a life information determination unit 15 and a learning unit 16 implemented by software. The determination device 100 according to the first embodiment includes pressure state information in the form of the relationship information shown in table 1. The determination device 100 according to the second embodiment is different in that the first relationship information shown in table 2 and the second relationship information in the form including a function and the like shown in fig. 10 are stored in the storage unit 8 and have pressure state information. The same applies otherwise. Hereinafter, the description will be omitted in the same manner as in the first embodiment.
TABLE 2
[ example 2 ]
The second relationship information shown in fig. 10 includes: the pressure input degree (vertical axis) of the subject input in the past and the intersection (icon of black solid circle mark) passing through the number (horizontal axis); a function F obtained by approximating the graph by a least square method with a specific regression equation (linear function in the case of fig. 10); and range information of pressure intensity (from level S1 to level S5) corresponding to the value of the function F. The degree of pressure input will be described later. The original values of the icons of fig. 10 are shown in table 3. Note that the second relationship information is not limited to the regression equation based on the least square method, and may be constructed by a learning method that reduces an error using training data.
TABLE 3
The subject No.
Job title
Number of passes
Degree of pressure input
1
Nurse (after class)
0
C1
2
Nurse (after class)
0
C1
3
Nurse (after class)
0
C1
4
Nurse (after class)
0
C1
5
Nurse (after class)
1
C1
6
Nurse (after class)
2
C1
7
Nurse (after class)
3
C1
8
Nurse (after class)
3
C1
9
Nurse (night shift)
8
C2
10
Nurse in ward
14
C3
11
Ward manager
17
C4
12
Pharmacy for ward
27
C5
The pressure input level in the present embodiment is a classification of the magnitude of pressure received by the subject, and is an external factor of the pressure assumed from the living information of the subject. In the present embodiment, 5 kinds of jobs of a nurse (after shift), a ward nurse manager, a ward manager, and a ward pharmacist are classified into stress input degrees (C1 to C5) as external factors of stress. The life information in the present embodiment is a factor that causes stress, such as the job title, the type of business, the effect on the business, the work time zone, and the work time of the subject. For example, nurses who are responsible for more patients are more responsible and stressed. Likewise, the physical burden on night shift workers is greater than that of white shift workers. In this case, the life information is the job, and the degree of stress input is the amount of stress assumed from the outside according to the job.
The living information determination unit 15 shown in fig. 9 is a functional unit that determines the degree of pressure input based on living information of the subject input from the input unit 91 or the like. The life information determination unit 15 reads the first relation information shown in table 2 from the storage unit 8 and determines the pressure input degree by referring to the read information. For example, when the post of the subject is a ward manager as the life information, the life information determination unit 15 determines the pressure input degree as the level of S4.
The comparison unit 13 of the determination unit 11 substitutes the number of passes counted by the counting unit 12 into the function F (see fig. 10) of the pressure state information read from the storage unit 8 to determine the pressure intensity level corresponding to the number of passes. For example, when the number of passes counted by the counting unit 12 is 21, the function value is about 4.2. Since this value is included in the range of the S4 level, the comparison unit 13 determines the pressure intensity level as the S4 level. Similarly, when the number of passages is 6, the comparison unit 13 determines the pressure intensity level as the level of S2. In this way, the determination unit 11 determines the pressure of the subject.
The learning unit 16 is a functional unit that learns pressure state information in which the degree of judgment of the degree of pressure input and the degree of judgment of the pressure state match each other to a higher degree. The learning unit 16 includes: a coincidence degree determination unit 17 for determining a coincidence degree between the determination result of the pressure input degree and the determination result of the pressure state; and an updating unit 18 for updating the second relationship information based on the matching degree. The learning unit 16 learns the pressure state information having a higher degree of matching with the pressure state determination result by repeating the update of the updating unit 18.
The coincidence degree determination unit 17 is a functional unit that determines the coincidence degree between the determination result of the pressure input degree determined by the living information determination unit 15 and the determination result of the pressure state determined by the determination unit 11. For example, if the determination result of the degree of pressure input is the same as the determination result of the pressure state, it is determined to be coincident. Otherwise, it is determined to be inconsistent.
The update unit 18 is a functional unit that updates the second relationship information based on the degree of matching determined by the degree of matching determination unit 17. When the matching degree determination unit 17 determines that the information pieces match, the update unit 18 updates the second relationship information. At the time of this update, the update unit 18 adds the number of passes of the determined subject and the degree of pressure input to the second relationship information, and updates the function F. By this update, the probability that the coincidence degree determination unit 17 determines coincidence in the subsequent pressure determination increases. That is, in the subsequent pressure determination, the degree of coincidence between the determination result of the pressure state and the determination result of the degree of pressure input becomes higher.
When the matching degree determination unit 17 determines that the information pieces do not match, the update unit 18 also updates the second relationship information. The update content is preferably weighted when the matching degree determination unit 17 determines that the contents match. For example, when the second relation information is approximated by the least square method, the weight of the inconsistent data can be reduced with respect to the consistent data. By this update, the probability that the coincidence degree determination unit 17 determines coincidence in the subsequent pressure determination increases. That is, in the subsequent pressure determination, the degree of coincidence between the determination result of the pressure state and the determination result of the degree of pressure input becomes higher.
The updating unit 18 issues a command to the determination unit 11 to perform the second determination based on the new function F after updating the second relationship information. The control unit 10 acquires the second determination result from the determination unit 11 and outputs the result to the output unit 92. The second determination result becomes a statistically more reliable value as the number of data increases.
The updating unit 18 repeats the above updating every time the pressure determination is performed. This improves the degree of matching between the determination result of the pressure state and the determination result of the degree of pressure input every time the determination device 100 performs pressure determination, and enables pressure determination reflecting the actual pressure state.
[ third embodiment ]
The determination unit 11 of the determination device 100 according to the first embodiment can be used for the following purposes: in order to assist the doctor in diagnosis and treatment of a disease, the determination unit 11 determines (identifies) the virus type contained in the sample L1, and identifies a disease of the subject based on the determined virus type. That is, the determination device 100 according to the third embodiment can be used as a virus detection device. Hereinafter, the structure, operation, and operational effects different from those of the first embodiment will be mainly described. Fig. 12 illustrates a configuration of a determination device 100 according to a third embodiment.
The determination unit 11 includes a disease determination unit 14 in addition to the counting unit 12 and the comparison unit 13 (an example of a virus determination unit).
In the first embodiment, the comparison unit 13 compares the passage number counted by the counting unit 12 with the pressure state information read from the storage unit 8 to determine the pressure intensity level corresponding to the passage number. Instead, the comparison unit 13 of the third embodiment determines the type of virus that generates the peak Pk based on the peak Pk corresponding to the trigger transmitted from the counting unit 12 and waveform information described later, as described later.
In the first embodiment, the counting unit 12 detects the number of peaks Pk in the waveform acquired in a specific time period while the voltage is applied to the first electrode 72, and counts the number of passes by considering the number of peaks Pk as the number of passes. Instead, the counting unit 12 of the third embodiment determines the presence of the peak Pk in the waveform acquired within a specific time period in a state where the voltage is applied to the first electrode 72, counts the number of the peaks, and when the presence of the peak Pk is identified, issues a command (hereinafter referred to as a trigger) for determining the type of the virus to the comparison unit 13 as described later, and then counts the type and the number of the viruses determined by the comparison unit 13. The comparison unit 13 completes the judgment of the virus type within 10 minutes at the latest as described later. The determination result of the comparing unit 13 is output to the output unit 92.
The disease determination unit 14 determines the disease of the subject based on the type of virus determined by the comparison unit 13, the number of each type of virus, and disease information described later. Details of the disease determination will be described later. The determination result of the disease determination unit 14 is output to the output unit 92. The judgment of a disease and its concept of the present embodiment include not only the judgment of a disease name (disease name) but also the judgment of information (e.g., information on viral activity) necessary for determining a treatment course (e.g., a prescription for administration).
In the storage unit 8, waveform information of a peak Pk (see fig. 5 and the like) corresponding to a known virus and disease information corresponding to a known virus are stored instead of or in addition to the pressure state information.
The waveform information is a database containing information on the shape of the peak Pk corresponding to a known virus. The waveform information includes information that can determine which kind of virus each peak Pk of the waveform acquired from the detection unit 7 is by comparison with each peak Pk of the waveform acquired from the detection unit 7.
The shape (waveform) of the peak Pk is illustrated in fig. 13 to 15. The waveform of fig. 13 is one peak Pk as a whole, which is an example of a case where there are a plurality of sub-peaks Pka (peak division) separately observable therein. Although two sub-peaks Pka having different wave heights and pulse widths are shown in fig. 13, the wave heights and pulse widths of the sub-peaks Pka may be substantially equal. The waveform of fig. 14 is an example of a broad-peak-shaped peak Pk, particularly in the case of a tailing peak (tailing). Examples other than the peak Pk having a broad peak shape include a leading peak (leading), although not shown. The waveform of fig. 15 illustrates a case where a shoulder peak (shoulder peak) Pkb is present in a part of the peak Pk. There are cases where a plurality of shoulder peaks Pkb occur.
The waveform information of the present embodiment is a learning model (an example of a learned model, hereinafter simply referred to as a learned model) constructed by learning in advance by machine learning such as deep learning technology. The waveform information of the present embodiment is as follows: a plurality of known viruses having different types, and activities are prepared in advance, and the peak Pk of a certain virus (e.g., an active herpes simplex virus 1 type) when passing through the through-hole 6 is repeatedly acquired as training data and tuned (enhanced), so that the type of the virus can be determined with a sufficiently high probability (e.g., a probability of 99% or more) when the peak Pk of the virus is input. In other words, when the learned model is used, if the comparing unit 13 acquires one pulse, the type of virus can be determined with a sufficiently high probability.
When the waveform information is constructed by learning, the learning can be performed focusing on the feature amount of the peak Pk. As the feature amount, for example, the above-described pulse wave height Ip, pulse wave width td, trailing peak, leading edge peak, shoulder peak Pkb, or the like may be used, and the number of sub-peaks Pka, wave height of each sub-peak Pka, and pulse wave width may be used. Further, when the waveform information is constructed by learning, the feature may be extracted by folding or merging the shapes of the peaks Pk, thereby being used for learning.
The disease information is a database containing information for identifying a disease from the type and the number of counts of viruses. The disease information includes information capable of identifying a disease based on the kind of virus and the passage number of each kind. For example, table 4 illustrates a case where, particularly when a virus is active, information for identifying a disease based on the number of types and models of each virus is used as disease information.
For example, in table 4, if the number of passes of the active herpes simplex virus type 1 (HSV-1) is equal to or greater than the threshold value α (e.g., 30), the disease determination section 14 determines that the administration of the antiviral drug is effective as HSV keratoherpes virus. Since the herpes simplex virus 1 type is counted (detected) from the body fluid depending on the stress state even in a healthy person, it may not be determined as a viral disease as long as the number of passes is less than the threshold value a.
In table 4, if the number of active Varicella Zoster Virus (VZV) passes is equal to or greater than the threshold value β (for example, 1), the disease determination section 14 determines that the administration of the antiviral drug is effective VZV uveitis. Further, if the number of active Human Cytomegaloviruses (HCMVs) passing through is equal to or greater than a threshold value ζ (e.g., 1), the disease determination unit 14 determines that the administration of the antiviral drug is effective as CMV keratodermitis. Since these viruses are not counted from the body fluid in healthy persons, even if one virus is detected, the disease is identified.
TABLE 4
Species of virus
HSV-1
VZV
…
HCMV
Threshold of passing number
α
β
ζ
Disease and disorder
HSV corneal herpes
VZV uveitis
CMV corneal endophthalmitis
In the present embodiment, the process of determining the type of virus contained in the body fluid of the subject is realized by causing the control unit 10(CPU) to execute a virus determination program (hereinafter, may be simply referred to as a program) for realizing the determination. The program causes the control unit 10 to acquire waveform information (an example of reception processing) from the detection unit 7, causes the determination unit 11 of the control unit 10 to apply the acquired waveform information to the learned model (an example of virus determination processing), and causes the control unit 10 to transmit the determination result to the output unit 92 or the like (an example of transmission processing). The program may be stored in the storage section 8. In the case of storing the program in the storage unit 8, the storage unit 8 may be stored in a non-transitory storage medium (for example, a magnetic storage medium, an optical disk, a flash memory, or the like).
[ procedure for Virus detection and disease assessment ]
The flow of virus detection will be described. Hereinafter, a case in which the storage unit 8 includes the disease information shown in table 4 will be described as an example. The measurer or the like stores the sample L1 in the sample storage section 2. Then, the electrolyte L2 was stored in the electrolyte storage section 3. Then, the sample L1 is brought into contact with the electrolyte L2 through the through hole 6 to be in an electrically conductive state (contact step).
When a measurer or the like inputs measurement conditions such as a measurement time (a time period during which a voltage is applied and continuously counted) and a detection start command to the control unit 10 via the input unit 91 after the sample L1 and the electrolyte L2 are brought into an electrically conductive state, the control unit 10 gives a command to the power supply unit 71 to apply a voltage under a condition corresponding to the measurement conditions. In response to this instruction, the power supply section 71 applies a voltage to the first electrode 72 under a specific condition.
The measurement time in the present embodiment is set to 5 minutes in advance. As described above, in the case of using the learned model, if the comparing unit 13 acquires one pulse, the type of virus can be determined with sufficiently high probability, and when the virus content (number concentration of viruses) of the sample L1 is small, it is necessary to continue the application of voltage and the like until one virus is counted (detected). If the measurement time is set to, for example, 1 minute or more and 10 minutes or less, preferably 1 minute or more and 5 minutes or less, the determination of the virus can be performed quickly and accurately.
In general, when a virus cannot be counted even if voltage application is continued for a certain time or more, the subject is healthy. Therefore, in the present embodiment, the measurement time of 5 minutes is preset as a reference of the measurement time sufficient for determining whether or not the patient is healthy. In order to further improve the accuracy of the judgment of health, the measurement time may be set with 10 minutes as an upper limit. Although the measurement time may be set to 10 minutes or more, it should be noted that the diagnosis by the doctor may be delayed or the waiting time of the patient may be extended.
If the measurement time is set too short, it may be erroneously determined that no virus is present (erroneously determined to be healthy) when the virus content is small. It is therefore advisable to ensure a measurement time of at least 10 seconds or more.
When a specific voltage is applied to the first electrode 72, the viruses of the sample L1 move toward the second electrode 73 by electrophoresis. When the virus passes through the through hole 6 during the movement, a peak Pk is generated in the waveform of the ion current (current detection step, passage step). An example of a waveform obtained from sample L1 of a patient with HSV corneal herpes is shown in figure 16. An example of a waveform obtained from sample L1 of a patient with CMV keratitis is shown in fig. 17. In fig. 16 and 17, a plurality of peaks Pk are generated.
The counting section 12 identifies the generation of the peaks Pk in the waveform, counts the number of identified peaks Pk as the number of passes, and sends one trigger per peak Pk to the comparing section 13.
When the comparison unit 13 receives the trigger, the peak Pk corresponding to the trigger is applied to the learned model read from the storage unit 8, and the virus type in which the peak Pk has occurred is determined (virus determination step). When the comparing unit 13 determines the virus type, information including the virus type is transmitted to the counting unit 12. The counting unit 12 that has received the information including the virus type counts the number of viruses for each virus type.
The disease determination unit 14 identifies a disease based on the type of virus determined by the comparison unit 13, the number of viruses counted for each type of virus by the counting unit 12, and disease information including information shown in table 4. If the disease determination unit 14 determines the type of virus and counts the number of viruses of each type, the disease can be immediately (e.g., simultaneously) identified.
For example, if there are 19 HSV corneal herpes, 15 varicella/zoster virus (VZV) and 0 person cytomegalovirus (HCMV), VZV uveitis judged to be effective for administration of antiviral drugs is considered a disease.
For example, if there are 32 HSV corneal herpes, 0 varicella/zoster virus (VZV) and 0 Human Cytomegalovirus (HCMV), HSV corneal herpes judged to be effective for administration of antiviral drugs is considered a disease.
For example, if there are 21 HSV corneal herpes, 0 varicella/zoster virus (VZV) and 0 Human Cytomegalovirus (HCMV), then no disease is identified. In this case, as in the first embodiment, a strong stress can be identified instead of the identification of a disease.
[ fourth embodiment ]
As shown in fig. 18, the fourth embodiment shows a case where the determination device 100 used as a virus detection device is configured by the terminal CL, the determination server SV, and the storage unit 8 which is a cloud server. The determination server SV and the terminal CL can be connected in bidirectional communication via the internet W (network N). Fig. 18 shows a case where a plurality of (2 in fig. 18) terminals CL are connected to the determination server SV so as to be capable of two-way communication. The following description deals with differences from the third embodiment.
The determination server SV has a control unit 10. Although fig. 18 illustrates a case where the determination server SV can access various information (including a learned model and a virus determination program) stored in the storage unit 8 via the network N, the determination server SV may have the storage unit 8.
The terminal CL includes the inspection board 1, and a terminal-side control unit 10a, a detection unit 7, a power supply unit 71, an input unit 91, and an output unit 92, each of which is capable of bidirectional communication connection by an internal bus D (network N).
The terminal-side control unit 10a communicates with the control unit 10 (determination server SV) via the internet W. The terminal-side control unit 10a controls operations of the inspection substrate 1, the detection unit 7, the power supply unit 71, the input unit 91, and the output unit 92 based on instructions from the control unit 10 in addition to input of instructions and the like from the input unit 10.
In the present embodiment, the process of determining the type of virus contained in the body fluid of the subject is realized by causing the determination server SV (control unit 10) to execute a virus determination program for realizing the determination. The program causes the control unit 10 to acquire waveform information (an example of reception processing) from the terminal CL, causes the determination unit 11 of the determination server SV to apply the acquired waveform information to the learned model (an example of virus determination processing), and causes the determination server SV to transmit the determination result of the determination unit 11 to the terminal CL that has acquired the waveform information (an example of transmission processing).
The terminal CL executes a terminal program that transmits waveform information to the determination server SV (control unit 10), receives the determination result from the determination server SV, and outputs the determination result to the output unit 92 or the like.
An example of the operation flow of this program is shown in fig. 19. Hereinafter, the determination device 100 will be described with reference to fig. 18, and the operation flow of the determination device 100 will be described with reference to fig. 19.
When waveform information is acquired in the terminal CL based on an input instruction or the like for starting measurement from the input unit 91 and the waveform information is transmitted to the determination server SV (#01), the determination server SV (control unit 10) receives the waveform information (# 11).
When the comparison unit 13 of the determination unit 11 inputs the received waveform information to the learned model (#12) and identifies the virus type with a probability equal to or higher than a specific probability (for example, equal to or higher than 99%) (#13, yes), the control unit 10 transmits the identified virus type to the terminal CL as the determination result (# 14). In the terminal CL, the terminal-side control unit 10a receives the determination result transmitted from the determination server SV, outputs the determination result (#02) from the output unit 92, and ends the determination. The output determination result is referred to by a doctor or the like.
After the determination result is transmitted to the terminal CL (#14), if the update of the learned model is permitted in the initial setting of the determination server SV (#15, yes), the learned model is updated based on the received waveform information to end the determination. If the update of the learned model is not permitted in the initial setting of the determination server SV (#16, No), the determination is ended directly. Note that steps #15 and #16 may be omitted.
When the type of virus is not determined with a probability equal to or higher than the specific probability (#13, No) and the peak Pk is not detected (#17, No), the control unit 10 transmits the determination result that No virus is detected to the terminal CL (# 14).
When the type of virus is not determined with a probability equal to or higher than a specific probability (#13, No) and the peak Pk (#17, Yes) is detected, waveform information is stored for the purpose of accumulating cases in the future (#18), and the determination is ended. Note that step #18 may be omitted.
Thus, the determination device 100 can also be realized by network computing such as a client server or a cloud server.
[ modified example of the embodiment ]
[ modification 1 ]
In the above embodiment, the case where the tear liquid is used as the body fluid of the subject has been described, in which the tear liquid is used as the cleansing liquid containing the tear liquid collected by cleansing the conjunctival sac of the subject with a phosphate buffered saline (physiological saline). In this case, the physiological saline solution for washing the conjunctival sac of the subject may contain an additive other than a buffer (pH adjuster) such as salt (sodium chloride) or sodium hydrogen phosphate.
Examples of the additive in this case include L-cysteine, homocysteine, glutathione, homocysteine, cysteine persulfate, glutathione persulfate, homocysteine persulfate, glucose, calcium chloride, magnesium chloride, and potassium chloride.
When glutathione is contained as an additive to a physiological saline solution, the physiological saline solution can be prepared to have a liquid property (hydrogen ion index) of about Ph7.1 to 8.1 and an osmotic pressure ratio of about 1.0 to 1.1. For example, the liquid properties and osmotic pressure ratio can be adjusted by blending sodium chloride, magnesium chloride, potassium chloride, sodium hydrogen phosphate, sodium hydrogen carbonate, sodium citrate, sodium acetate, and the like.
Particularly, when glutathione is contained as an additive to a physiological saline solution, the accuracy of virus type determination of the virus is improved, and therefore, this is preferable. This is due to: by including glutathione in the saline solution, the activity of the virus is improved, or the activity of the virus is prevented from being lowered to improve the storage stability of the sample, and the shape of the peak Pk of the virus depending on the virus species can be detected more significantly.
An example in which the accuracy of determination of the virus type of a virus is improved will be described. First, an anterior aqueous humor (an example of an aqueous humor) is collected as a body fluid from an eyeball of a patient with CMV iritis.
Next, a sample (sample A) in which the collected anterior chamber water was stored at 4 ℃ for 2 hours directly, a sample (sample B) in which the anterior chamber water was stored at 4 ℃ for 2 hours after being diluted 2-fold with a phosphate-buffered physiological saline, and a sample (sample C) in which the anterior chamber water was stored at 4 ℃ for 2 hours after being diluted 2-fold with a commercially available intraocular perfusate (product name: BSS Plus 500 intraocular perfusate 0.0184%, manufacturer: Nippon Archon) containing glutathione as an active ingredient were prepared. Sample C corresponds to the case where sample L1 of the present embodiment contains glutathione.
The reuse judgment apparatus 100 detected the virus in the sample A, B, C. The number of human cytomegaloviruses detected in A, B, C samples containing aqueous humor obtained from the same patient (detection time: 5 minutes) was 0, 7, and 12, respectively.
From the results of the detection of human cytomegalovirus from the above sample A, B, C, it can be seen that: when sample L1 contains glutathione, the accuracy of virus detection is improved, and the accuracy of virus type determination is improved.
[ modification 2 ]
In the above embodiment, a case where an electrolyte solution such as a physiological saline solution is used as the electrolyte solution L2 has been described. However, the electrolyte L2 may contain additives in addition to common salt (sodium chloride).
Examples of the additive in this case include L-cysteine, homocysteine, glutathione, homocysteine, cysteine persulfate, glutathione persulfate, homocysteine persulfate, glucose, calcium chloride, magnesium chloride, potassium chloride, and other pH regulators. Examples of the pH adjuster include sodium hydrogen phosphate, sodium hydrogen carbonate, sodium citrate, sodium acetate, and lactic acid.
When glutathione is contained as an additive in the electrolyte solution L2, the electrolyte solution L2 can be prepared to have a liquid property (hydrogen ion index) of about ph7.1 to 8.1 and an osmotic pressure ratio of about 1.0 to 1.1. For example, the liquid properties and osmotic pressure ratio can be adjusted by blending sodium chloride, magnesium chloride, potassium chloride, sodium hydrogen phosphate, sodium hydrogen carbonate, sodium citrate, sodium acetate, and the like.
Particularly, when glutathione is contained as an additive to electrolyte L2, it is preferable because the accuracy of virus type determination of the virus is improved. This is due to: when glutathione is contained in the electrolyte L2, the activity of the virus is improved, or the activity of the virus is prevented from being lowered to improve the storage stability of the sample, whereby the shape of the peak Pk of the virus depending on the virus species can be detected more significantly.
[ example ]
[ clinical diagnosis, judgment or examination result for each case ]
Hereinafter, the determination result of the virus type by the determination device 100 according to the third embodiment will be described by comparing it with the clinical diagnosis result of a doctor and the determination result of the virus type by the real-time PCR method (hereinafter, simply referred to as the PCR method).
The results of clinical diagnosis of cases 1 to 9, the type of sample, the results of determination by the determination device 100, and the results of detection by the PCR method are shown in table 4. The sample determined by the determination device 100 is the same as the sample subjected to the PCR method. In the clinical diagnosis of cases 1 to 9, the doctor did not use the judgment result obtained by the judgment apparatus 100 and the detection result obtained by the PCR method.
The sample supplied to the determination device 100 was collected from the subject, diluted 2-fold with a commercially available intraocular perfusate (product name: BSS Plus 500 intraocular perfusate 0.0184%, manufacturer: Nippon Archon) containing glutathione as an active ingredient, and used. The determination device 100 sets the measurement time to 5 minutes and outputs (detects) the virus species having the active virus. The measurement time of the PCR method was set to about 3 hours, and the virus species were detected. Here, the presence or absence of viral activity could not be determined by the PCR method.
TABLE 5
Species of virus
HSV-1
VZV
…
HCMV
Threshold of passing number
α
β
ζ
Disease and disorder
HSV corneal herpes
VZV uveitis
CMV corneal endophthalmitis
In case 1 and cases 3 to 6, the virus corresponding to the clinical diagnosis result was detected by the determination device 100, but the virus was not detected by the PCR method and was determined to be negative. In cases 2, 7, and 8, viruses corresponding to clinical diagnosis were detected by the determination device 100 and the PCR method. In conclusion, the following is considered: the determination device 100 has high correlation with clinical diagnosis and higher sensitivity of determination or detection than the PCR method.
In case 9, CMV iritis due to infection with HCMV was diagnosed in clinical diagnosis, and HCMV was detected by the PCR method. However, no active virus was detected by the determination device 100. In case 9, since the clinical diagnosis result shows no antiviral effect, it is considered that in case 9, active virus is not present in the sample (in the subject) at the time of sampling the sample. In addition, it is considered that in case 9, the inactive virus leaked out into the tear fluid and was detected by the PCR method.
It is difficult for a doctor who is responsible for recovering the health of a subject to make a judgment of excluding an effective treatment (for example, administration of an antiviral drug) from treatment options when it is diagnosed that there is no active virus in the subject or when there is an active virus in the subject as in case 9 when a specific symptom or disease name (CMV iritis in this case) can be clinically diagnosed. However, the doctor can select a more appropriate diagnosis and treatment method by obtaining information that no active virus is present in the subject based on the determination result of the determination device 100.
As described above, according to cases 1 to 9, it is considered that: the doctor can make a more appropriate diagnosis based on the detection result of the active virus by the determination device 100.
As described above, it is possible to provide a simple, quick and accurate virus detection method, a virus detection device for realizing the virus detection method, a virus determination program, a pressure determination method, a pressure determination device for realizing the pressure determination method, and a determination device for realizing the pressure determination device.
[ other embodiments ]
(1) In the above embodiment, the herpes simplex virus 1 (HSV-1) was described as the second microparticle, but the microparticle is not limited to the herpes simplex virus 1 (HSV-1). As the fine particles, there may be used various types of herpesviruses such as Human Cytomegalovirus (HCMV) and human herpesvirus 6 (HHV-6), and viruses (virus types) other than herpesviruses such as non-enveloped adenovirus like herpesviruses.
The pulse wave height Ip and pulse wave width td of the peak Pk for three different virus species, HSV-1, HCMV and HHV-6, are shown in FIG. 11. As shown in fig. 11, the shape of the peak differs for each virus type, but even when the virus type differs, the peak Pk can be detected. Therefore, even when the types of viruses are different, the number of passes can be counted based on the waveform.
(2) In the above embodiment, the case where the determination unit 11 has the counting unit 12 that counts the number of passes based on the waveform and the comparison unit 13 that compares the number of passes with the pressure state information, and determines the pressure state of the subject based on the number of passes and the pressure state information has been described, but the present invention is not limited to this.
For example, the determination unit 11 may further detect the shape of the peak Pk such as the pulse wave height Ip and the pulse wave width td by the counting unit 12 or the like, and determine the type of the virus corresponding to each peak Pk. Then, the passage number and the distribution of the virus species in the passage number are acquired, and the pressure state of the subject can be determined by comparing them with the pressure state information. Further, without determining the type of virus, the number of passes and information on the distribution of the shapes of the peaks Pk, such as the pulse wave height Ip and the pulse wave width td among the number of passes, may be acquired and compared with the pressure state information to determine the pressure state of the subject.
In this way, when the pressure state of the subject is determined by acquiring the distribution of the virus types or acquiring the information on the distribution of the shapes of the peaks Pk, the pressure state information includes at least the relationship information between the number of each virus type of the sample and the pressure intensity, or the relationship information between the information on the number of the peaks Pk and the distribution of the shapes of the peaks Pk and the pressure intensity, which are acquired in advance under the specific measurement conditions, as the pressure state information.
The pressure intensity level information that can be included in the relationship information when the pressure state of the subject is determined by acquiring the distribution of the virus species or acquiring the information on the distribution of the peak Pk shape as described above is explained. For example, it may contain information on the level of stress intensity such that if the distribution amount of human herpesvirus type 6 (HHV-6) is large, it is subjected to a strong long-term stress rather than a short-term stress. Based on such relationship information, the determination unit 11 can determine that a strong long-term pressure is applied when the distribution amount of the HHV-6 is large, and can determine that a strong short-term pressure is applied when the distribution amount of the HHV-6 is small.
(3) In the above embodiment, the case where the body fluid is a tear fluid has been described, but the body fluid is not limited to a tear fluid. For example, other body fluids such as aqueous humor (also simply referred to as aqueous humor), vitreous humor, saliva, sweat, urine, and the like can be used.
(4) In the above-described embodiment, the case where the term replaced with the term that can intuitively grasp the pressure state is read in accordance with the pressure intensity level is exemplified, but the representation is not limited to this embodiment. For example, a job representation that is replaced to correspond to a stress intensity level may be read. The display may be provided so that the user who uses the determination device 100 to make the determination or the subject who is determined by the determination device 100 can easily grasp the type and size of the external factors such as the responsibility and the operation time. In the present embodiment, for example, the case of dealing with the job of a nurse or the like working in a hospital or the like is cited.
(5) In the above-described embodiment, the external stress factor for classifying the degree of stress input is exemplified by a job such as a nurse working in a hospital or the like, but the external factor for stress is not limited thereto. For example, commute time, work period, etc. may be used, or these factors may be added together for use.
(6) In the above embodiment, the inner surface 6a of the through-hole 6 is surface-treated to have a negative surface charge having the same polarity as that of the virus, but surface modification is not essential and may not be performed.
(7) In the above embodiment, the case where the disease information includes information that the disease determination unit 14 can identify a disease based on the type of virus and the number of passage of each virus has been described. However, the disease information is not limited thereto, and may include other information. For example, the disease information may include information that the disease determination unit 14 can determine one disease or a plurality of highly probable diseases from the combination of virus types and the amount (number) of each virus type. The case of identifying a plurality of diseases based on the combination of the types of viruses and the amount (number) of each virus type may include information that the disease determination unit 14 can identify the diseases in a proper order of probability (for example, probability).
(8) In the first to third embodiments, the case where the determination device 100 includes the inspection substrate 1 as an inspection chip, the detection unit 7 that acquires a waveform from the inspection substrate 1, the control unit 10 such as a CPU that determines the pressure state of the subject based on information acquired from the inspection substrate 1, and the power supply unit 71, and the detection unit 7 and the like are connected to the control unit 10 via the network N has been described. However, the configuration of the determination device 100 is not limited to this case.
The determination device 100 may be, for example, a portable article in which a determination device main body (not shown) is a unit in which the control unit 10, the detection unit 7, and the network N are integrated. The inspection substrate 1 can be an article that can be arbitrarily connected to or provided in the determination device main body and an inspection chip that can be disconnected from or removed from the determination device main body, and that is highly portable.
In this case, by further setting the test board 1 to be freely attachable to and detachable from the determination device main body, the body fluid is collected near the patient and stored in the test board 1, and convenience can be improved by only allowing the test board 1 in which the body fluid is stored to be easily moved (transported). In addition, by preparing a plurality of test substrates 1 for the determination device 100, and making them as exclusive articles for each patient or disposable articles, it is possible to prevent cross-infection.
(9) In the fourth embodiment, the case where the terminal CL includes the terminal-side control unit 10a, the detection unit 7, the power supply unit 71, the input unit 91, and the output unit 92, which are bidirectionally communicably connected to the inspection board 1 via the internal bus D (network N), respectively, has been described. However, the configuration of the terminal CL is not limited to this case.
The terminal CL may be, for example, a portable article in which the determination device main body (not shown) is a unit in which the terminal-side control unit 10a, the detection unit 7, the power supply unit 71, and the internal bus D are integrated. The inspection substrate 1 can be an article that can be arbitrarily connected to or provided in the determination device main body and an inspection chip that can be disconnected from or removed from the determination device main body, and that is highly portable.
In this case, by further setting the test board 1 to be freely attachable to and detachable from the determination device main body, the body fluid is collected near the patient and stored in the test board 1, and convenience can be improved by only allowing the test board 1 in which the body fluid is stored to be easily moved (transported). In addition, by preparing a plurality of test substrates 1 for the determination device 100 and making them as exclusive articles for each patient or disposable articles, it is possible to prevent cross-contamination and improve the determination accuracy.
(10) In the above-described embodiment, the herpes simplex virus type 1 (HSV-1(HHV1)), the varicella/herpes zoster virus (VZV (HHV3)), the human cytomegalovirus (HCMV (HHV5)), and the human herpes virus type 6 (HHV-6) have been exemplified as the virus (particularly, herpes virus), but the virus that can be determined by the determination device 100 is not limited thereto. The determination device 100 can determine the types of various herpesviruses including at least human herpesvirus HHV 1-8. The determination device 100 can also determine the type of a virus other than a herpesvirus, for example, adenovirus, enterovirus, Coxsackie virus (Coxsackie virus), human papilloma virus (human papillomavirus), rubella virus, and other clinically important viruses.
(11) In the third embodiment, a learning model capable of determining the type of a certain virus (for example, active herpes simplex virus 1 type) when the peak Pk of the virus is input is exemplified as the learned model (waveform information). However, the types of viruses that can be learned by the learned model are not limited to active viruses, and include inactive viruses. By learning the peak Pk including the active virus and the peak Pk including the inactive virus using the learned model, the comparing section 13 can easily determine the presence or absence of activity as the type of virus. Thus, the disease determination section 14 can identify a disease in consideration of the presence or absence of viral activity.
(12) In the description of the virus detection flow of the third embodiment, the case where the storage unit 8 contains, as the disease information, information for identifying a disease based on the number of passage of each virus type or model, particularly when a virus is active, and the disease determination unit 14 determines that the administration of an antiviral drug is effective, as shown in table 4, has been described as an example. However, the disease information stored in the storage unit 8 may include information for identifying a disease by differentiating the type, model, and presence or absence of activity of a virus. In this case, even if the disease is the same (for example, VZV uveitis), the disease determination section 14 may make a determination to distinguish between a disease in which the administration of the antiviral drug is effective and a disease in which the administration of the antiviral drug is ineffective.
For example, when active viruses and inactive viruses are counted, particularly when the number of active viruses counted is large, the disease determination unit 14 determines that the administration of the antiviral drug is effective.
For example, the disease determination unit 14 determines a disease in which the administration of the antiviral drug is ineffective when only inactive viruses are counted, or when most (for example, 80%) of the total number of passage of the counted viruses are inactive viruses although active viruses and inactive viruses are counted. A disease in which administration of an antiviral drug is ineffective is, for example, a state in which it has been clinically diagnosed as a disease caused by a virus but is already in a healing process (viral clearance is being accomplished) by an immunological action in a patient's body, and means that it does not contribute to treatment and quality of life (QOL) of the patient even if the antiviral drug has been administered.
Note that, unless a contradiction occurs, the structure disclosed in the above embodiment (including other embodiments, the same applies hereinafter) may be applied in combination with the structure disclosed in other embodiments. In addition, the embodiments disclosed in the present specification are examples, and the embodiments of the present invention are not limited thereto, and may be modified as appropriate within a range not departing from the object of the present invention.
Industrial applicability
The present invention can be applied to a simple, quick and accurate virus detection method, virus detection device, pressure determination program, pressure determination method, and pressure determination device.
Description of the symbols
2: sample storage part
3: electrolyte storage part
5: partition wall
6: thru hole (thru hole portion)
7: detection part
8: storage unit
10: control unit
10 a: terminal side control unit
11: determination unit (pressure determination unit)
12: counting part
13: comparison part (virus determination part)
14: disease determination unit
15: life information determination unit
16: learning part
17: coincidence degree determination unit
18: updating part
70: current meter
91: input unit
100: determination device (pressure determination device, virus detection device)
L1: sample(s)
L2: electrolyte solution
V: virus
- 上一篇：一种医用注射器针头装配设备
- 下一篇：凝血系统分析装置