阅读说明：本技术 凝血系统分析装置和方法 (Coagulation system analysis device and method ) 是由 林义人 李丞玟 渕上彩 町田贤三 于 2019-09-06 设计创作，主要内容包括：提供了一种凝血系统分析装置,包括：一对电极；施加单元,其被配置为以预定的时间间隔向该对电极施加交流电压；测量单元,其被配置为测量放置在该对电极之间的血液样本的复介电常数；以及分析单元,其被配置为通过使用至少两种或多种类型的凝血相关测定,基于在作用于血液样本的抗凝血作用被解除之后的时间间隔测量的预定时间段内的特定频率的复介电常数来分析凝血因子的活性和凝血抑制因子的活性。(Provided is a coagulation system analysis device including: a pair of electrodes; an applying unit configured to apply an alternating voltage to the pair of electrodes at predetermined time intervals; a measurement unit configured to measure a complex permittivity of a blood sample placed between the pair of electrodes; and an analysis unit configured to analyze the activity of the coagulation factor and the activity of the coagulation inhibiting factor based on a complex permittivity of a specific frequency within a predetermined period of time measured at a time interval after an anticoagulation effect acting on the blood sample is released, by using at least two or more types of coagulation-related assays.)

1. A coagulation system analysis device comprising:

circuitry configured to:

performing a first coagulation-related assay on a first blood sample;

performing a second coagulation-related assay on a second blood sample; and is

Assessing the activity of a coagulation factor and the activity of a coagulation inhibitor based at least in part on the output of the first coagulation related assay and the output of the second coagulation related assay,

wherein the first blood sample and the second blood sample are dispensed from the same blood sample collected from a patient.

2. The coagulation system analysis device of claim 1, wherein the circuit is further configured to output information indicative of a risk of thrombosis of the patient and/or information indicative of a risk of bleeding of the patient based at least in part on the output of the first coagulation-related assay and the output of the second coagulation-related assay.

3. The coagulation system analysis device of claim 1, wherein the first coagulation-related assay comprises an extrinsic coagulation pathway trigger assay and the second coagulation-related assay comprises an intrinsic coagulation pathway trigger assay.

4. The coagulation system analysis device of claim 3, wherein the extrinsic coagulation pathway trigger assay comprises an extrinsic weak trigger assay and/or the intrinsic coagulation pathway trigger assay comprises an intrinsic weak trigger assay.

5. The coagulation system analysis device of claim 1, wherein the first coagulation-related assay and the second coagulation-related assay are selected from the group consisting of an assay that does not trigger a coagulation reaction, a strong endogenous trigger assay, a weak endogenous trigger assay, a strong exogenous trigger assay, and a weak exogenous trigger assay.

6. The coagulation system analysis device of claim 1, wherein the first coagulation-related assay comprises a strong trigger assay and the second coagulation-related assay comprises a weak trigger assay.

7. The coagulation system analysis device of claim 1, wherein the output of the first coagulation-related assay and/or the output of the second coagulation-related assay comprises a coagulation time.

8. The coagulation system analysis device of claim 1, wherein the first coagulation-related assay comprises an intrinsic weak trigger assay configured to output a first coagulation time, the second coagulation-related assay comprises an intrinsic strong trigger assay configured to output a second coagulation time, and the circuit is further configured to perform a third coagulation-related assay comprising an extrinsic weak trigger assay configured to output a third coagulation time.

9. The coagulation system analysis device according to claim 8, wherein determining the activity of the coagulation factor and the activity of the coagulation inhibitor comprises:

(i) comparing the first clotting time to a first threshold;

(ii) comparing the second clotting time to a second threshold;

(iii) comparing the third clotting time to a third threshold; and is

Outputting information indicative of a risk of thrombosis of the patient and/or information indicative of a risk of bleeding of the patient based at least in part on one or more of comparison (i), comparison (ii), and comparison (iii).

10. The coagulation system analysis device of claim 1, wherein the circuit is further configured to:

determining a first permittivity spectrum based on the output of the first coagulation-related assay;

extracting values of one or more features from the first permittivity spectrum;

determining a second permittivity spectrum based on the output of the second coagulation related assay;

extracting values of the one or more features from the second permittivity spectrum; and is

Evaluating the activity of the coagulation factor and the activity of the coagulation inhibiting factor based on the values of the one or more features extracted from the first permittivity spectrum and the values of the one or more features extracted from the second permittivity spectrum.

11. The coagulation system analysis device of claim 10, wherein the one or more characteristics comprise: the time CT at which the dielectric constant has a minimum value and the time CT3 at which the slope of the dielectric constant is maximum.

12. The coagulation system analysis device of claim 1, wherein the circuit is further configured to:

providing information associated with the output of the first coagulation-related assay and information associated with the output of the second coagulation-related assay as inputs to a training model, and wherein the activity of the coagulation factor in the blood sample and the activity of the coagulation inhibiting factor in the blood sample are based at least in part on an output of the training model.

13. The coagulation system analysis device according to claim 12,

the information associated with the output of the first coagulation-related assay comprises values of one or more features extracted from a permittivity spectrum determined based on the output of the first coagulation-related assay; and is

The information associated with the output of the second coagulation-related assay includes values of the one or more features extracted from a permittivity spectrum determined based on the output of the second coagulation-related assay.

14. The coagulation system analysis device of claim 12, wherein the trained models comprise models trained using one or more machine learning techniques.

15. The coagulation system analysis device of claim 1, wherein the circuit further comprises:

a measurement unit configured to perform the first coagulation-related assay and the second coagulation-related assay, the measurement unit being configured to measure a dielectric impedance of the first blood sample and the second blood sample when each blood sample is arranged between electrodes to which an alternating voltage is applied.

16. A method of analyzing a blood sample to determine an amount of risk of thrombosis and/or an amount of risk of bleeding in a patient associated with the blood sample, the method comprising:

performing a first coagulation-related assay on a first blood sample;

performing a second coagulation-related assay on a second blood sample; and is

Assessing the activity of a coagulation factor and the activity of a coagulation inhibitor based at least in part on the output of the first coagulation related assay and the output of the second coagulation related assay,

wherein the first blood sample and the second blood sample are dispensed from the same blood sample collected from a patient.

17. A coagulation analysis system comprising:

a first electrode and a second electrode arranged opposite to the first electrode such that a container comprising a first blood sample can be arranged between the first electrode and the second electrode;

a voltage generator configured to apply an alternating voltage to the first electrode and the second electrode; and

circuitry configured to:

performing a first coagulation-related assay on the first blood sample while the alternating voltage is applied to the first electrode and the second electrode;

performing a second coagulation-related assay on a second blood sample; and is

Assessing the activity of a coagulation factor and the activity of a coagulation inhibitor based at least in part on the output of the first coagulation related assay and the output of the second coagulation related assay,

wherein the first blood sample and the second blood sample are dispensed from the same blood sample collected from a patient.

Technical Field

The present technology relates to a coagulation system analysis device, a coagulation system analysis method, and a coagulation system analysis program.

Background

In the past, coagulation tests have existed as a clinically performed method of analyzing blood conditions. Coagulation tests represented by a prothrombin time-international normalized ratio (PT-INR) and Activated Partial Thromboplastin Time (APTT) are known as general coagulation tests. In these methods, proteins involved in the coagulation reaction are analyzed for their coagulation reactivity. Such proteins are contained in plasma obtained by centrifuging a blood sample.

The above test methods are used for functional tests for assessing the ability to extrinsic and intrinsic coagulation. In these tests, substances triggering the extrinsic coagulation reaction and the intrinsic coagulation reaction are added in excess to obtain test results in a short time. These tests were performed using plasma obtained by centrifuging a blood sample. In such a test, since cellular components (such as platelets and red blood cells) that play an important role in the coagulation reaction in vivo are removed by centrifugation, a difference occurs between the test result and the actual clinical condition in many cases.

Note that examples of another functional test include thromboelastography and thromboelastometry, which are commercialized as TEG (registered trademark) and ROTEM (registered trademark), respectively. These functional tests are not extensive enough because, for example, (1) the measurement is not automated and the test results depend on the skill of the measurer, (2) it is susceptible to vibration, (3) the Quality Control (QC) procedure is complex and QC reagents are expensive, and (4) skill is required to obtain an interpretation of the output signal (thromboelastogram). Furthermore, since sensitivity to lack or inhibition of extrinsic and intrinsic coagulation factors is not sufficiently shown, there is a possibility that the demand in the medical field cannot be satisfied.

In addition, in the above-mentioned conventional functional test, a coagulation initiator is generally used as a test reagent in an excess amount larger than the in vivo reaction. These tests are therefore suitable for assessing a significant decrease in the coagulation capacity, i.e. the bleeding tendency, but not for assessing a significant increase in the coagulation capacity, i.e. the thrombogenic tendency or slight changes in the coagulation capacity. Note that, in the thromboelastometry, there is an assay in which measurement is performed by newly adding calcium without using a coagulation initiator that artificially activates the extrinsic coagulation pathway or the intrinsic coagulation pathway. In this assay, fibrin gel formed at an early stage of a coagulation reaction is broken due to rotational displacement during measurement, and thus it is difficult to perform correct measurement. Specifically, in such an assay, there are the following problems: for samples with low fibrinogen or platelet content, reproducibility of the measurement cannot be expected.

In recent years, as another method capable of evaluating a coagulation measurement simply and accurately, a method of performing a dielectric measurement of a coagulation process has been proposed (for example, see japanese patent application publication No. 2010-181400 and japanese patent application publication No. 2012-194087). In this method, a blood sample is placed in a capacitor-like sample portion including a pair of electrodes and the like, and an alternating electric field is applied to the sample portion to measure a change in complex permittivity that occurs during coagulation of the blood sample. Y. hayashi et al, Analytical Chemistry 87(19), 10072-. Furthermore, i.uchimura et al, biorhology 53,209-219(2016) showed that by using this method, the increase in the blood coagulation ability can be evaluated with high sensitivity, which has been difficult to evaluate by another method. However, this method also has the following problems: it is difficult to correctly assess complex coagulation kinetics.

Reference list

Patent document

Patent document 1: japanese patent application publication No. 2010-181400,

patent document 2: japanese patent application laid-open No. 2012-194087.

Non-patent document

Non-patent document 1: y. hayashi et al, Analytical Chemistry 87(19),10072-10079(2015),

non-patent document 2: uchimura et al, Biorhology 53,209-219 (2016).

Disclosure of Invention

Technical problem

As described above, it is difficult to correctly evaluate complex coagulation kinetics using existing methods.

In this respect, it is desirable to provide a blood coagulation system analyzer (blood-clotting-system analysis apparatus) capable of evaluating coagulation kinetics with high accuracy.

Solution to the problem

According to the present disclosure, a blood coagulation system analysis device (blood coagulation system analysis device) is provided. The coagulation system analysis device includes circuitry configured to perform a first coagulation-related assay on a first blood sample; performing a second coagulation-related assay on a second blood sample; and assessing the activity of the coagulation factor and the activity of the coagulation inhibiting factor based at least in part on the output of the first coagulation-related assay and the output of the second coagulation-related assay, wherein the first blood sample and the second blood sample are dispensed from the same blood sample collected from the patient.

In accordance with the present disclosure, a method of analyzing a blood sample to determine an amount of risk of thrombosis and/or an amount of risk of bleeding in a patient associated with the blood sample is provided. The method comprises performing a first coagulation-related assay on a first blood sample; performing a second coagulation-related assay on a second blood sample; and assessing the activity of the coagulation factor and the activity of the coagulation inhibiting factor based at least in part on the output of the first coagulation-related assay and the output of the second coagulation-related assay, wherein the first blood sample and the second blood sample are dispensed from the same blood sample collected from the patient.

In accordance with the present disclosure, a coagulation assay system is provided. The coagulation analysis system includes: a first electrode and a second electrode arranged opposite to the first electrode such that a container comprising a first blood sample may be arranged between the first electrode and the second electrode; a voltage generator configured to apply an alternating voltage to the first electrode and the second electrode; and a circuit. The circuit is configured to perform a first coagulation-related assay on a first blood sample when an alternating voltage is applied to the first electrode and the second electrode; performing a second coagulation-related assay on a second blood sample; and assessing the activity of the coagulation factor and the activity of the coagulation inhibiting factor based at least in part on the output of the first coagulation-related assay and the output of the second coagulation-related assay, wherein the first blood sample and the second blood sample are dispensed from the same blood sample collected from the patient.

Drawings

Fig. 1 is a schematic conceptual diagram schematically showing an example of the concept of a coagulation system analysis device 100 according to an embodiment of the present technology.

Fig. 2 is a sectional view schematically showing an example of an electrical measurement container 101 according to an embodiment of the present technology.

Fig. 3 is a drawing alternative diagram describing an example of measurement of a complex permittivity spectrum (three-dimensional).

Fig. 4 is a drawing alternative diagram describing an example of measurement of a complex permittivity spectrum (two-dimensional).

Fig. 5 is a drawing alternative diagram describing an example of feature quantities extracted from a complex permittivity spectrum.

FIG. 6 FIGS. 6A to 6D are each a diagram alternative showing the screening of the change in the coagulation test and the coagulation fibrinolytic factor with the timing of blood collection.

FIG. 7 FIGS. 7E to 7H are each a diagram alternative showing the screening of the change in the coagulation test and the coagulation fibrinolytic factor with the timing of blood collection.

Fig. 8A and 8B are each a diagram alternative diagram showing a variation of the ROTEM measurement result with the blood collection timing.

Fig. 9C to 9E are each a diagram alternative diagram showing a variation of the ROTEM measurement result with the blood collection timing.

Fig. 10A to 10D are each a diagram alternative diagram showing a variation of a DBCM measurement analysis result with a blood collection timing.

Fig. 11E to 11H are each a diagram alternative diagram showing a variation of a DBCM measurement analysis result with a blood collection timing.

Fig. 12 is a block diagram showing an example of a coagulation analysis method in view of thorough finding of a thrombosis risk.

Fig. 13 is a block diagram illustrating an example of a coagulation analysis method for determining a bleeding risk using an exogenous weak trigger assay and an endogenous weak trigger assay.

Detailed Description

In the following, advantageous embodiments of the present technique will be described with reference to the drawings.

The embodiments described below illustrate examples of typical embodiments of the present technology, and the scope of the present technology is not narrowly construed by the embodiments. Note that description will be made in the following order.

1. Blood coagulation system analysis device 100

(1) A pair of electrodes 1a and 1b

(1-1) Electrical measuring Container 101

(1-2) connection unit 102

(1-3) Container holding Unit 103

(2) Application unit 2

(3) Measuring cell 3

(4) Analysis unit 4

(5) Output unit 5

(6) Display unit 6

(7) Memory cell 7

(8) Measurement condition control unit 8

(9) Temperature control unit 9

(10) Blood sample supply unit 10

(11) Reagent supply unit 11

(12) Accuracy management unit 12

(13) Drive mechanism 13

(14) Sample Standby Unit 14

(15) Stirring mechanism 15

(16) User interface 16

(17) Server 17

(18) Others

2. Blood coagulation analysis method

(1) Applying step

(2) Measurement procedure

(3) Analytical procedure

1. Blood coagulation system analysis device 100

The blood coagulation system analysis device 100 includes at least a pair of electrodes 1a and 1b, an application unit 2, a measurement unit 3, and an analysis unit 4. Further, the blood coagulation system analysis device 100 may include other units such as an output unit 5, a display unit 6, a storage unit 7, a measurement condition control unit 8, a temperature control unit 9, a blood sample supply unit 10, a reagent supply unit 11, a quality control unit 12, a drive mechanism 13, a sample standby unit 14, an agitation mechanism 15, a user interface 16, and a server 17 as necessary.

Hereinafter, the details thereof will be described.

(1) A pair of electrodes 1a and 1b

The pair of electrodes 1a and 1B is brought into contact with the blood sample B at the time of measurement, and applies a necessary voltage to the blood sample B.

The arrangement, form, and the like of the pair of electrodes 1a and 1B are not particularly limited, and may be freely designed as needed as long as it can apply a necessary voltage to the blood sample B. In the present technique, it is advantageous that a pair of electrodes 1a and 1b are integrally formed in an electrical measurement container 101 described below.

Also, the material forming the electrodes 1a and 1b is not particularly limited. One or two or more known conductive materials can be freely selected and used as needed as long as the condition or the like of the blood sample B to be analyzed is not affected. Specifically, examples of such materials include titanium, aluminum, stainless steel, platinum, gold, copper, and graphite.

In the present technique, it is advantageous to form the electrodes 1a and 1b from a conductive material containing titanium among these materials. Titanium is suitable for performing measurements of blood sample B, due to its low clotting activity properties relative to blood samples.

(1-1) Electrical measuring Container 101

Fig. 2 is a cross-sectional view schematically illustrating an example of an electrical measurement container 101 in accordance with embodiments of the present technique. The electrical measurement container 101 holds a blood sample B to be analyzed. In the blood coagulation system analysis device 100 according to the embodiment of the present technology, the number of the electrical measurement containers 101 is not particularly limited, and one or more electrical measurement containers 101 may be freely arranged as appropriate according to the amount, type, and the like of the blood sample B to be analyzed.

In the blood coagulation system analysis device 100 according to the embodiment of the present technology, the measurement of the complex permittivity is performed while the electrical measurement container 101 holds the blood sample B. Therefore, it is advantageous that the electrical measurement container 101 is configured to be sealable while holding the blood sample B. However, the electrical measurement container 101 does not have to be configured to be airtight as long as it can be stationary in the time it takes to measure the complex dielectric constant and the configuration does not affect the measurement.

The specific method and sealing method for introducing the blood sample B into the electrical measurement container 101 are not particularly limited. The blood sample B may be introduced by a free method as appropriate depending on the form of the electrical measurement container 101 and the like. Examples of such a method include a method of providing a lid in the electrical measurement container 101 and closing the lid to seal after introducing the blood sample B using a pipette or the like.

The form of the electrical measurement container 101 is not particularly limited, and may be freely designed as appropriate as long as the blood sample B to be analyzed can be held in the device. Further, electrical measurement container 101 may include one or more containers.

The specific form of the electrical measurement container 101 is not particularly limited, and may be freely designed as appropriate according to the condition of the blood sample B or the like, as long as the electrical measurement container 101 can hold the blood sample B to be analyzed. Examples of such forms include a cylinder, a polygonal cylinder having a polygonal cross section (triangular, square, or larger), a cone, a polygonal cone having a polygonal cross section (triangular, square, or larger), and a form obtained by combining one or two or more of them.

Further, the material forming the container 101 is also not particularly limited, and may be appropriately freely selected as long as the condition or the like of the blood sample B to be analyzed is not affected. In particular, in the present technology, it is advantageous that the container 101 is formed of a resin from the viewpoint of ease of processing and forming, and the like. In the present technology, the type and the like of the resin that can be used are not particularly limited. One or two or more kinds of resins that can be used for holding the blood sample B can be freely selected and used as appropriate. Examples of such resins include hydrophobic and insulating polymers such as polypropylene, polymethylmethacrylate, polystyrene, acrylic, polysulfone, and polytetrafluoroethylene, copolymers and polymer blends.

Specifically, in the present technique, it is advantageous that the electrical measurement container 101 is formed of one or more resins selected from polypropylene, polystyrene, acrylic acid, and polysulfone. These resins are suitable for performing measurements on blood samples due to their low clotting activity relative to blood samples.

Note that in the present technique, a well-known disposable cartridge type included may also be used as the electrical measurement container 101.

In the present technique, it is advantageous to provide a plurality of electrical measurement containers 101 that include two or more types of coagulation related assays described below. Therefore, the analysis of the magnitude of the thrombus formation risk can be performed efficiently in the analysis unit 4 described below. Further, in the present technology, it is advantageous that reagents constituting two or more types of coagulation-related assays are packaged in each of the plurality of electrical measurement containers 101 in advance for each assay.

In the present technique, in the case of using a reagent, as described above, a predetermined reagent may be held in advance in a solidified state or as it is in a liquid state in the electrical measurement container 101. For example, an anticoagulant, a coagulation initiator, an intrinsic coagulation pathway initiator, an extrinsic coagulation pathway initiator, a calcium salt, and the like may be placed in the container 101 in advance. By making the container 101 hold the reagent in advance as described above, the reagent supply unit 11 or a part holding the reagent described below is not required, which makes it possible to miniaturize the apparatus and reduce the cost. Further, since the user does not need to replace the reagent and does not need to maintain devices such as the reagent supply unit 11 and the portion holding the reagent, usability can be improved.

(1-2) connection unit 102

The connection unit 102 electrically connects the application unit 2 described below with the electrodes 1a and 1 b. The specific form of the connection unit 102 is not particularly limited, and may be freely designed as appropriate as long as it can electrically connect the application unit 2 with the electrodes 1a and 1 b.

(1-3) Container holding Unit 103

The container holding unit 103 holds the electrical measurement container 101. The specific form of the container holding unit 103 is not particularly limited, and may be freely designed as appropriate as long as it can hold the container 101 containing the blood sample B to be analyzed.

Further, the material forming the container holding unit 103 is also not particularly limited, and may be freely selected as appropriate depending on the form of the electrical measurement container 101 and the like.

Further, in the present technology, the container holding unit 103 may have a function (for example, a barcode reader or the like) of automatically reading information related to the container 101 from an information recording medium provided in the electrical measurement container 101. Examples of the information recording medium include an IC card, an IC tag, a card having a barcode or a matrix 2D code, or a paper or sticker on which a barcode or a matrix 2D code is printed.

(2) Application unit 2

The applying unit 2 applies an alternating voltage to the pair of electrodes 1a and 1b at predetermined intervals. More specifically, for example, the application unit 2 applies an alternating voltage to the pair of electrodes 1a and 1b at a start time of a time point at which an instruction to start measurement is received or a time point at which the power supply of the apparatus 100 is turned on. More specifically, the applying unit 2 applies an alternating voltage of a set frequency or a frequency controlled by the measurement condition control unit 8 described later to the pair of electrodes 1a and 1b for each set measurement interval or a measurement interval controlled by the measurement condition control unit 8 described later.

(3) Measuring cell 3

The measurement unit 3 measures the complex permittivity of the blood sample placed between the pair of electrodes 1a and 1 b. The configuration of the measurement unit 3 may be freely designed as appropriate as long as the measurement unit 3 can measure the complex permittivity of the blood sample B to be measured. Specifically, an impedance analyzer, a network analyzer, or the like may be employed as the measurement unit 3.

More specifically, for example, the measurement unit 3 may be configured to chronologically measure the impedance of the blood sample B obtained by applying an alternating voltage to the blood sample B by the application unit 2, and chronologically measure the impedance of the blood sample B between the electrodes 1a and 1B at a start time of a time point at which an instruction to start measurement is received or a time point at which the power of the apparatus 100 is turned on. The complex permittivity is then derived from the measured impedance. To derive the complex permittivity, a known function or relationship showing the relationship between impedance and permittivity may be used.

The measurement result of the measurement unit 3 may be obtained as a three-dimensional complex permittivity spectrum (fig. 3) having frequency, time, and permittivity as coordinate axes, or a two-dimensional complex permittivity spectrum (fig. 4) having two of frequency, time, and permittivity as coordinate axes. The Z-axis in fig. 3 indicates the real part of the complex permittivity at each time and each frequency.

Fig. 4 corresponds to a two-dimensional spectrum obtained by cutting the three-dimensional spectrum shown in fig. 3 at a frequency of 760 kHz. Symbol (a) in fig. 4 indicates a peak associated with the formation of a red blood cell stack (red blood cells), and symbol (B) indicates a peak associated with the blood sample coagulation process. The inventors of the present invention disclosed in japanese patent application publication No. 2010-181400 that the time change of the permittivity of the blood sample reflects the coagulation process of the blood sample. Therefore, the complex permittivity spectrum obtained by the measurement unit 3 is an index quantitatively indicating the blood coagulation ability of the blood sample. Based on the change of such index, information on the coagulation ability of the blood sample, such as coagulation time, coagulation rate, and coagulation intensity, can be acquired.

(4) Analysis unit 4

The analysis unit 4 analyzes the activity of the coagulation factor and the activity of the coagulation inhibiting factor using at least two or more types of coagulation-related assays, based on the complex permittivity of a specific frequency within a predetermined period of time measured at the above-described time interval starting at the time after the anticoagulation action on the blood sample B is released. Note that in this specification, the concept of "activity" includes physiological activity including a decrease in the reaction rate.

As described above, it becomes apparent in examining the present technology that it is difficult to correctly assess complex coagulation kinetics with existing methods. Specifically, as shown in example 1 described below, it has been found that in the case where (i) the blood coagulation ability is partially reduced due to the deficiency of a coagulation factor, but high coagulability is observed relative to other factors and the like, or (ii) the cause of high coagulability is due to a decrease in the actual activity of a coagulation inhibitory factor (particularly, antithrombin) under physiological conditions, the risk of thrombus formation cannot be correctly analyzed with the existing methods.

The thrombosis risk in the case of (i) above is a thrombosis risk that is found during recovery after surgery using cardiopulmonary bypass (cardiorespiratory bypass) and cannot be found by normal tests. Further, in the case of (ii) above, if a vitamin K antagonist inhibitor (e.g., warfarin) which is generally prescribed for preventing thrombosis is used, the amount of antithrombin or the like which is a coagulation inhibitory factor is also reduced. Also in this case, the coagulation test such as the PT-INR test cannot reflect the decrease in the amount of antithrombin due to the characteristics of the test, which results in a test result of coagulation prolongation, and erroneously determines that the risk of thrombosis has decreased. In practice, however, both the ability to clot and the ability to inhibit clotting (primarily by antithrombin) are reduced under physiological conditions, and the balance between both clotting ability and ability to inhibit clotting is important for the risk of thrombosis. Therefore, it is not necessarily said that the risk of thrombosis is reduced. To prevent this misunderstanding, it is necessary to perform not only the coagulation test but also the antithrombin activity measurement.

However, even if antithrombin activity measurement is performed, the following cannot be dealt with. Specifically, the case where the amount (concentration) of antithrombin is not reduced but the physiological (dynamic) activity is decreased is problematic. In other words, a case where the amount of antithrombin is not changed but the reaction rate of antithrombin is decreased cannot be dealt with. The existing antithrombin activity test or quantitative test cannot reflect the slight change of the reaction rate of antithrombin. Meanwhile, in the case where whether or not thrombus is generated in vivo is a problem, not only the amount of antithrombin but also the reaction rate of antithrombin is important because the reaction of accelerating the activity of the coagulation factor for thrombin generation competes with the reaction of inhibiting thrombin generation. This thrombotic risk in relation to the reaction rate of antithrombin has not been widely recognized so far.

Meanwhile, in particular, the blood coagulation system analysis device according to the embodiment of the present technology can find even a sample having the above-described risk of thrombus formation without exception. In particular, the risk of thrombosis, which has not been recognized so far, can also be evaluated, and the coagulation kinetics can be evaluated with high accuracy. Thus, customized therapy matching the coagulation status of the patient can be performed in all diseases where post-operative thrombosis hemostasis control or coagulation abnormalities are a problem.

Further, under the condition where the coagulation factor reduction and the coagulation inhibitory factor activity reduction coexist, although it is necessary to perform not only the coagulation test but also the measurement of the blood coagulation factor and the like to evaluate both risks in the case of the existing method, both risks can be easily evaluated by the present technology and the treatment regimen can be decided based on the risk determination result. Further, by applying the present technology to general medical examinations, it is possible to know the risk before thrombosis, cerebral hemorrhage, or the like occurs.

In the present technology, it is advantageous that the two or more types of coagulation-related assays described above are selected from the group consisting of an assay that does not trigger a coagulation reaction, an intrinsic coagulation pathway triggering assay (hereinafter also referred to as "strong intrinsic trigger assay"), an assay that triggers the intrinsic coagulation pathway more weakly than the intrinsic coagulation pathway triggering assay (hereinafter also referred to as "weak intrinsic trigger assay"), an extrinsic coagulation pathway triggering assay (hereinafter also referred to as "strong extrinsic trigger assay"), and an assay that triggers the extrinsic coagulation pathway more weakly than the extrinsic coagulation pathway triggering assay (hereinafter also referred to as "weak extrinsic trigger assay").

Further, in the present technology, it is advantageous that the two or more types of coagulation-related assays comprise an endogenous weak trigger assay and/or an exogenous weak trigger assay.

The strong endogenous trigger assay includes, for example, calcium salts to address the anticoagulant effect of citric acid, and an endogenous coagulation pathway initiator at a concentration that strongly triggers the endogenous coagulation pathway. Examples of intrinsic coagulation pathway initiators include ellagic acid. In this case, for example, in general APTT test reagents, those using ellagic acid as a coagulation activator (e.g., actin SFL) can be used. Regarding the final concentration of ellagic acid, blood samples: the ratio of APTT test reagents is preferably 18: 1 to 200: 1, and more preferably 50: 1 to 150: 1, and the optimal final concentration is 90: 1.

weak intrinsic trigger assays include, for example, calcium salts to address the anticoagulant effect of citric acid, and coagulation pathway initiators at concentrations that weakly trigger the coagulation pathway. Examples of intrinsic coagulation pathway initiators include ellagic acid. In this case, as regards the final concentration of ellagic acid, the blood sample: the ratio of APTT test reagents is preferably 230: 1 to 2300: 1, and more preferably 450: 1 to 1800: 1, and the optimal final concentration is 900: 1.

the above-mentioned extrinsic strong trigger assay includes, for example, a calcium salt for solving the anticoagulation effect of citric acid, and an extrinsic coagulation pathway initiator at a concentration that strongly triggers the extrinsic coagulation pathway. Examples of extrinsic coagulation pathway initiators include Tissue Factor (TF). In this case, the final concentration of tissue factor is preferably higher than 5pM, and more preferably not lower than 10pM, and the optimum final concentration is 50 pM.

The above exogenous weak triggering assay includes, for example, calcium salts to address the anticoagulant effect of citric acid, and an exogenous coagulation pathway initiator at a concentration that weakly triggers the exogenous coagulation pathway. Examples of extrinsic coagulation pathway initiators include tissue factor. In this case, the final concentration of the tissue factor is preferably not more than 5pM, and more preferably 0.2pM to 2.0pM, and the optimum final concentration is 0.6pM to 0.7 pM.

In the present technology, the blood coagulation factor whose activity is to be analyzed by the analysis unit 4 is not particularly limited, but may be specifically an intrinsic blood coagulation factor as shown in example 2 described below. Further, the coagulation inhibitory factor whose activity is to be analyzed by the analyzing unit 4 is also not particularly limited, but may be specifically antithrombin as shown in example 2 described below.

More specifically, the analysis unit 4 extracts a feature quantity of a complex permittivity spectrum of a specific frequency measured using a certain measurement, and determines whether the feature quantity exceeds a determination reference set in advance, for example, based on the measurement results of a specific number of healthy persons and/or patients. For example, a threshold value may be simply set as a determination reference. However, it is advantageous to use a value defined based on a function of the clotting time(s) and the clotting time (w). Clotting time(s) was measured using a strong trigger assay and clotting time (w) was measured using a weak trigger assay.

Similarly, the analysis unit 4 extracts the feature quantity of the complex permittivity spectrum of a specific frequency measured using another measurement different from the above-described specific measurement, and determines whether or not the feature quantity exceeds a determination reference.

Then, the analysis unit 4 compares the determination result obtained by using a specific assay with the determination result obtained by using a different assay, and performs classification based on the determination result to finally determine the condition of each sample (each blood sample), i.e., whether the activity of the coagulation factor and the activity of the coagulation inhibitory factor are normal or decreased.

Note that as the above-described characteristic amount, a time index relating to a blood sample coagulation reaction, an index relating to a reaction rate, or the like can be employed. Further, in the present technology, a new feature amount or value may be calculated by combining a feature amount by a specific measurement and a feature amount by a different measurement, and the new feature amount or value may be compared with a determination reference set in advance.

Fig. 5 is a drawing alternative diagram describing an example of feature quantities extracted from a complex permittivity spectrum. In fig. 5, the vertical axis indicates the dielectric constant, and the horizontal axis indicates time. The upper graph is based on measurements around a frequency of 1MHz (not less than 100kHz to less than 3 MHz). The lower graph is based on measurements near a 10MHz frequency (3MHz to 30 MHz).

In the present technology, as the above-described feature amount, a time feature amount and/or a slope feature amount extracted from a complex permittivity spectrum at a specific frequency can be used. Further, the slope characteristic amount may be extracted based on the time characteristic amount extracted from the complex permittivity spectrum at a specific frequency. More specifically, for example, one or more selected from the group consisting of: time CT0 giving the maximum value of the complex dielectric constant at a low frequency of 100kHz or more and less than 3 MHz; time CT1 (not shown) which gives the maximum slope at low frequencies; maximum slope CFR at low frequency; a time CT4 (not shown) when the absolute value of the slope reaches a predetermined ratio (preferably, 50%) of the maximum slope CFR after the time CT 1; time CT giving the minimum value of complex dielectric constant at high frequency of 3MHz to 30 MHz; time CT3 which gives the maximum slope at high frequency; maximum slope CFR2 at high frequency; time CT2 which gives the minimum value of the complex permittivity when a straight line is drawn from time CT3 with the slope of the maximum slope CFR2 after time CT and before time CT 3; and a time CT5 (not shown) when the absolute value of the slope reaches a predetermined ratio (preferably, 50%) of the maximum slope CFR2 after the time CT 3. Further, an operation value of these characteristic amounts, an operation value having a measured complex permittivity, or the like may also be used. In addition, the analysis unit 4 may also analyze the activity of the coagulation factor and the activity of the coagulation inhibitor by using a training model including a model trained using one or more machine learning techniques.

(5) Output unit 5

The output unit 5 outputs the analysis result obtained by the analysis unit 4. In the present technology, the configuration of the output unit 5 is not particularly limited. For example, the output unit 5 may be configured to generate a notification signal at a specific time and notify the user of the result in real time only in the case where an abnormality analysis result is obtained during measurement. With this configuration, since the analysis result is notified to the user only at a specific point in time when the abnormal analysis result has been determined, usability is improved.

Further, the method of notifying the user is also not particularly limited. For example, the user can receive a notification via the display unit 6, a display, a printer, a speaker, an illumination device, or the like, which will be described later. Further, for example, a device having a communication function of transmitting an electronic mail or the like for notifying that a notification signal has been generated to a mobile device such as a cellular phone and a smart phone may be used together with the output unit 5.

Further, in the present technology, for example, the output unit 5 may have a function of notifying: in the case where a plurality of electrical measurement containers 101 including the above-described two or more types of coagulation-related assays are not set in the apparatus 100, although the magnitude of the risk of analyzing thrombosis is input to the apparatus 100 in advance, the user is warned or the like to prompt the user to set the containers 101.

Further, in the present technology, the output unit 5 may output the magnitude of the thrombus formation risk and/or the hemorrhage risk based on the analysis result of the analysis unit 4. Therefore, by using the coagulation system analysis device according to the embodiment of the present technology, even a sample (blood sample) having both a thrombosis risk and a bleeding risk can be thoroughly found, resulting in early treatment.

Further, in the present technology, the output unit 5 may further output the cause of the thrombosis risk and/or the bleeding risk based on the analysis result of the analysis unit 4. Therefore, not only each risk can be determined but also the cause of the risk can be known, and therefore, the demand for a quick medical site can be satisfied.

Note that the risk of thrombosis may be due to, for example, any one or more selected from the group consisting of exposure of tissue factor to a blood sample, reduction in antithrombin amount or antithrombin response rate, and enhancement of the intrinsic coagulation pathway. Furthermore, the risk of bleeding may be due to, for example, a deficiency in extrinsic and/or intrinsic coagulation factors. In other words, in the present technology, the risk of thrombosis and the risk of bleeding may have multiple causes or only one cause.

(6) Display unit 6

The display unit 6 displays the analysis result of the analysis unit 4, data relating to the complex permittivity measured by the measurement unit 3, a notification result from the output unit 5, and the like. The configuration of the display unit 6 is not particularly limited. For example, a display, a printer, or the like may be employed as the display unit 6. Further, in the present technology, the display unit 6 does not necessarily need to be provided, and an external display device may be connected.

(7) Memory cell 7

The storage unit 7 stores the analysis result of the analysis unit 4, data relating to the complex permittivity measured by the measurement unit 3, a notification result from the output unit 5, and the like. The configuration of the memory cell 7 is not particularly limited. For example, a hard disk drive, a flash memory, an SSD (solid state drive), or the like may be employed as the storage unit 7. Further, in the present technology, the storage unit 7 does not necessarily need to be provided, and an external storage device may be connected.

Further, in the present technology, an operation program and the like of the coagulation system analysis device 100 may be stored in the storage unit 7.

(8) Measurement condition control unit 8

The measurement condition control unit 8 controls the measurement time and/or the measurement frequency and the like in the measurement unit 3. As a specific method of controlling the measurement time, the measurement interval may be controlled in accordance with the amount of data required for target analysis or the like, or the timing of completing the measurement may be controlled in a case where, for example, the measurement value has been substantially leveled.

Further, the measurement frequency may also be controlled according to the type of the blood sample B to be measured, the measurement value required for the target analysis, and the like. Examples of the method of controlling the measurement frequency include a method of changing the frequency of an alternating voltage to be applied between the electrodes 1a and 1b, and a method of superimposing a plurality of frequencies to measure impedance at a plurality of frequencies. Examples of the specific method include a method of arranging a plurality of single-frequency analyzers side by side, a method of scanning frequencies, a method of superimposing frequencies using a filter and extracting information of each frequency, and a method of performing measurement using a response to an impulse.

(9) Temperature control unit 9

The temperature control unit 9 controls the temperature in the electrical measurement container 101. In the coagulation system analysis device 100 according to the embodiment of the present technology, the temperature control unit 9 does not necessarily need to be provided. However, in order to keep the blood sample B to be analyzed under optimal measurement conditions, a temperature control unit 9 is preferably provided.

Further, in the case where the sample standby unit 14 is provided as will be described later, the temperature control unit 9 may control the temperature in the sample standby unit 14. Further, in the case where a reagent is put into the blood sample B at the time of measurement or before the measurement, a temperature control unit 9 may be provided to control the temperature of the reagent. In this case, the temperature control unit 9 may be provided for temperature control in the electrical measurement container 101, temperature control in the sample standby unit 14, and temperature control of the reagent. Alternatively, one temperature control unit 9 may perform all temperature controls.

The specific method of controlling the temperature is not particularly limited. However, for example, by providing the container holding unit 103 with a temperature adjusting function, it is possible to cause the container holding unit 103 to function as the temperature control unit 9.

(10) Blood sample supply unit 10

The blood sample supply unit 10 automatically supplies the blood sample B to the electrical measurement container 101. In the blood coagulation system analysis device 100 according to the embodiment of the present technology, the blood sample supply unit 10 does not necessarily need to be provided. However, by providing the blood sample supply unit 8, it is possible to automatically perform in each step of analyzing the coagulation system.

The specific method of supplying the blood sample B is not particularly limited. However, the electrical measurement container 101 may be automatically supplied with the blood sample B by using a pipette and a tip attached to the end of the pipette, for example. In this case, in order to prevent measurement errors and the like, it is advantageous that the tip is disposable. Further, it is also possible to automatically supply the electrical measurement container 101 from the reservoir of the blood sample B by using a pump or the like. Further, it is also possible to automatically supply the blood sample B to the electrical measurement container 101 by using a permanent nozzle or the like. In this case, in order to prevent measurement errors and the like from occurring, it is advantageous to provide the nozzle with a cleaning function.

Further, in the present technology, the blood sample supply unit 10 may include a function (a barcode reader or the like) for identifying and automatically reading the type or the like of the blood sample B as a sample.

(11) Reagent supply unit 11

The reagent supply unit 11 automatically supplies one or more reagents to the electrical measurement container 101. In the coagulation system analysis device 100 according to the embodiment of the present technology, the reagent supply unit 11 does not necessarily need to be provided. However, by providing the reagent supply unit 11, each step of analyzing the coagulation system can be automatically performed.

The specific method of supplying the reagent is not particularly limited, and a method similar to the above-described blood sample supply unit 10 may be used. In particular, it is advantageous to supply the reagent by using a method capable of supplying a predetermined amount of reagent without contacting the electrical measurement container 101. For example, in the case of a liquid reagent, the reagent may be discharged and supplied. More specifically, for example, the liquid reagent may be discharged and supplied to the container 101 by introducing the liquid reagent into a discharge pipe in advance and blowing pressurized air separately connected via a pipe connected to the discharge pipe into the pipe in a short time. At this time, by adjusting the air pressure and the valve opening/closing time, the amount of the liquid reagent to be discharged can also be adjusted.

Further, in addition to blowing air, the liquid reagent may be discharged and supplied to the container 101 by using vaporization of the liquid reagent itself or by heating air dissolved therein. At this time, it is also possible to adjust the amount of generated bubbles and adjust the amount of liquid reagent to be discharged by adjusting the voltage and application time applied to the vaporization chamber where the heating element and the like are placed.

Further, it is also possible to supply the liquid reagent to the container 101 by driving a movable unit provided in a pipe line without using air using a piezoelectric element (piezo element) or the like, and conveying the liquid reagent in an amount determined by the volume of the movable unit. Further, for example, the agent may also be supplied by using a so-called ink-jet method in which a liquid agent is made into fine droplets and sprayed directly onto a desired container 101.

Further, in the present technology, the reagent supply unit 11 may be provided with a stirring function, a temperature control function, a function for identifying and automatically reading, for example, the type of a reagent (e.g., a barcode reader), and the like.

(12) Accuracy management unit 12

The accuracy management unit 12 manages the accuracy of the measurement unit 3. In the coagulation system analysis device 100 according to the embodiment of the present technology, the quality control unit 12 does not necessarily need to be provided. However, by providing the accuracy management unit 12, the measurement accuracy of the measurement unit 3 can be improved.

The specific method of managing the accuracy is not particularly limited, and a known accuracy management method can be freely used as appropriate. Examples of such methods include a method of managing the accuracy of the measurement unit 3 by performing calibration of the measurement unit 3, such as a method of performing calibration of the measurement unit 3 by placing a metal plate or the like for short-circuiting in the apparatus 100 and short-circuiting the electrode and the metal plate before starting measurement, a method of bringing a calibration jig or the like into contact with the electrode, and a method of performing calibration of the measurement unit 3 by placing a metal plate or the like in the same container as the form of the container 101 in which the blood sample B is to be placed and short-circuiting the electrode and the metal plate before starting measurement.

Further, the present technology is not limited to the above-described method, and a free method, for example, a method of managing the accuracy of the measurement unit 3 by checking the state of the measurement unit 3 before actual measurement and calibrating the measurement unit 3 by performing the above-described calibration or the like only when there is an abnormality, may be appropriately selected and used.

(13) Drive mechanism 13

The drive mechanism 13 is used to move the electrical measuring container 101 in the measuring unit 3 according to various purposes. For example, by moving the container 110 to a direction that changes the direction of gravity applied to the blood sample B held in the container 110, the measurement value can be prevented from being affected by the sedimentation of the sedimentation component in the blood sample B.

Further, for example, the electric measuring container 101 may be driven so that the applying unit 2 and the electrodes 1a and 1b may be disconnected from each other at the time of non-measurement, and the applying unit 2 and the electrodes 1a and 1b may be electrically connected to each other at the time of measurement.

Further, for example, in the case where a plurality of electrical measurement containers 101 are provided, by configuring the containers 101 to be movable, measurement, blood sample supply, reagent supply, and the like can be performed by moving the container 110 to a necessary portion. That is, since it is not necessary to move the measurement unit 3, the blood sample supply unit 10, the reagent supply unit 11, and the like to the target electric measurement container 101, it is not necessary to provide a driving unit or the like for moving the respective units, and it is possible to miniaturize the apparatus and reduce the cost.

(14) Sample Standby Unit 14

The sample standby unit 14 makes the separated blood sample B stand by before measurement. In the coagulation system analysis device 100 according to the embodiment of the present technology, the sample standby unit 14 does not necessarily need to be provided. However, by providing the sample standby unit 14, the dielectric constant can be measured smoothly.

In the present technology, the sample standby unit 14 may be provided with a stirring function, a temperature control function, a mechanism for moving to the electrical measurement container 101, a function for identifying and automatically reading, for example, the type of the blood sample B (e.g., a barcode reader), an automatic opening function, and the like.

(15) Stirring mechanism 15

The stirring mechanism 15 stirs the blood sample B, and stirs the blood sample B and the reagent. In the coagulation system analysis device 100 according to the embodiment of the present technology, the stirring mechanism 15 does not necessarily need to be provided. However, for example, in the case where the blood sample B contains a sedimentary component, or in the case where a reagent is added to the blood sample B at the time of measurement, it is advantageous to provide the stirring mechanism 15.

The specific stirring method is not particularly limited, and a known stirring method can be freely used as appropriate. Examples of such methods include stirring by pipetting, stirring using a stirring rod, a stirring bar, or the like, and stirring by inverting a container containing the blood sample B or the reagent.

(16) User interface 16

The user interface 16 is a part for user operation. The user can access the respective units of the coagulation system analysis device 100 via the user interface 16.

(17) Server 17

The server 17 includes at least a storage unit that stores data acquired by the measurement unit 3 and/or analysis results acquired by the analysis unit 4, and is connected to at least the measurement unit 3 and/or the analysis unit 4 via a network.

Further, the server 17 is capable of managing various types of data uploaded from the respective units of the blood coagulation system analysis apparatus 100, and outputting various types of data to the display unit 6 or the like in response to an instruction from the user.

(18) Others

Note that the functions performed by the respective units of the blood coagulation system analysis device 100 according to the embodiment of the present technology may be stored as programs in a personal computer or hardware resources including a control unit including a CPU or the like and a recording medium (a nonvolatile memory (USB memory or the like), an HDD, a CD, or the like), and realized by the personal computer or the control unit.

2. Blood coagulation system analysis method

The coagulation system analysis method comprises at least an application step, a measurement step and an analysis step. In addition, the coagulation system analysis method may include other steps as necessary. Hereinafter, each step will be described in detail.

(1) Applying step

The applying step includes applying an alternating voltage to the pair of electrodes at predetermined time intervals. The detailed method is the same as the above-described method performed by the applying unit 2, and thus the description thereof is omitted here.

(2) Measurement procedure

The measuring step includes measuring a complex permittivity of a blood sample placed between a pair of electrodes. The detailed method is the same as the above-described method performed by the measurement unit 3, and thus the description thereof is omitted here.

(3) Analytical procedure

The analyzing step includes analyzing the activity of the coagulation factor and the activity of the coagulation inhibiting factor based on a complex permittivity of a specific frequency within a predetermined period of time measured at the above-described time interval after the anticoagulation action on the blood sample is released, by using at least two or more types of coagulation-related assays. The detailed method is the same as the above-described method performed by the analysis unit 4, and thus the description thereof is omitted here.

Note that a more specific analysis method is shown in fig. 12 of example 3 or fig. 13 of example 4 described below.

Examples of the invention

Hereinafter, the present technology will be described in more detail based on examples.

Note that the examples described below show examples of typical embodiments of the present invention, and the scope of the present technology is not narrowly construed by the examples.

< < example 1 >)

Blood was measured from 24 adult patients who were to undergo cardiovascular surgery using cardiopulmonary bypass.

< sample >

The timing of blood collection is as follows. Blood was collected into a blood collection tube containing citric acid as an anticoagulant.

(A) After induction of anesthesia and before the start of surgery

(B) Time to completion of neutralization of heparin with protamine following cardiopulmonary bypass

(C) Time to access ICU post-operatively

(D) One week after the operation

(E) One month after the operation

< measurement >

In addition to DBCM (dielectric coagulation assay) measurement, thromboelastometry, a test for screening coagulation, a quantitative test for coagulation factors, a thrombin generation test, and the like are performed.

For DBCM measurement, a dielectric coagulometer (prototype for experiment) (manufactured by sony corporation) was used. The measurement system comprises an automatic blood dispensing unit, a quadruple cassette holder controlled at 37 ℃ (within +0 ℃ or-1 ℃), an impedance analyzer board (frequency range: 100Hz to 40MHz) connected to a disposable cassette, and a computer. The cartridge is formed of polypropylene, and a pair of electrodes each formed of titanium is inserted therein. The cartridge is capable of measuring the complex permittivity of blood by acting as a parallel plate capacitor. Further, the pair of electrodes is arranged to be less prone to blood deposition. The cassette pre-packaged with reagents is arranged on the sample cassette holder. Note that the dielectric hemagglutination meter (prototype for experiment) used this time includes a quadruple cartridge holder, and four types of measurements can be simultaneously performed using different reagents.

The names of the reagents used in this example 1 and their corresponding assays are as follows: assay EX for activation of the extrinsic coagulation pathway by tissue factor; assay for activation of the intrinsic coagulation pathway by ellagic acid IN; assay PI in which the extrinsic coagulation pathway is activated by tissue factor in a state where platelet aggregation is inhibited by cytochalasin D; and L1, an assay for the activation of the extrinsic coagulation pathway by tissue factor in a state in which the fibrinolytic system is inhibited by aprotinin. Note that in addition to these agents, calcium chloride is often added to address the anticoagulant effect of citric acid. Furthermore, the sample was used as whole blood for measurement without special treatment.

Thromboelastometry measurements were performed using ROTEM δ (manufactured by TEM innovations GmBH) according to the procedure specified by the manufacturer. The assay used for the measurement is selected to correspond to the DBCM measurement. Specifically, the assay is an EXTEM assay in which the extrinsic coagulation pathway is activated by tissue factor, an INTEM assay in which the intrinsic coagulation pathway is activated by ellagic acid, a FIBTEM assay in which the extrinsic coagulation pathway is activated in a state in which platelet aggregation is inhibited by cytochalasin D, and an APTEM assay in which the extrinsic coagulation pathway is activated by tissue factor in a state in which the fibrinolytic system is inhibited by aprotinin. Note that the sample was used as whole blood for measurement without special treatment.

A test for screening coagulation and a quantitative test for coagulation factors were performed by a blood coagulation fibrinolysis measuring device (ACLTOP 300-CTS; manufactured by Instrument laboratories). It is necessary to use plasma from which blood cell components have been removed. For this, the blood sample was centrifuged at 3000rpm × 10min (at 20 ℃), the supernatant was recovered, and the supernatant was centrifuged at 3000rpm × 10min (at 20 ℃) to obtain a supernatant. The supernatant finally obtained was used. The plasma thus obtained is plasma from which substantially all platelets have been removed, and can be considered Platelet Poor Plasma (PPP). The measurement is performed using a program and a dedicated reagent specified by the manufacturer. The measurement items are shown in table 1 below.

[ Table 1]

[ measurement items of Table 1]

< results and discussion >

In the extrinsic coagulation tests, i.e., PT-INR (fig. 6A) and ROTEM EXTEM (fig. 8A), the clotting time after completion of cardiopulmonary bypass was prolonged compared to when anesthesia was induced prior to surgery, but the tendency to delay into the ICU after completion of surgery was alleviated. Of particular note is the prolonged clotting time of one week post-surgery and one month post-surgery as compared to the clotting time just prior to surgery. One reason for this is that FVII upstream of the extrinsic coagulation pathway is significantly reduced one week after surgery and one month after surgery (fig. 6D).

Focusing on the variation of each coagulation factor, many of these factors have lower values after completion of cardiopulmonary bypass (fig. 6C and 6D and 7E), and blood dilution by infusion also contributes to this. The trend differs depending on the measurement items for the factors of one week after surgery and one month after surgery. For example, fibrinogen has a high value especially one week after surgery due to post-operative inflammation. At the same time, it can be said that FVII has a considerably lower value than the trend of change of other factors (fig. 6D). The fibrinolysis indicators (i.e., DD (fig. 7F) and FDP (fig. 7G)) have very high values after completion of cardiopulmonary bypass and tend to be higher than values just before surgery and even one month after surgery. In addition, AT (fig. 7H) decreased with surgery but recovered one week after surgery.

Meanwhile, with respect to DBCM, the characteristic amount as the change in permittivity is 10MHz, and the results obtained by performing analysis and summarization for each measurement and blood collection timing with a time giving the minimum value as the time CT and with a time at which the slope with the increase in permittivity of coagulation is maximum (a time giving the maximum slope) as the time CT3 are shown in fig. 10A to 11H. Upon entering the ICU after completion of cardiopulmonary bypass, a generally prolonged trend is shown as compared to just prior to surgery. Such changes match the results of PT-INR or ROTEM EXTEM shown above.

However, after that (one week after surgery and one month after surgery) the time CT levels of each assay (EX, LI and PI) that triggered the extrinsic pathway of coagulation have returned to the level just before surgery and the time CT for some samples has decreased. From this finding, the risk of thrombosis (hypercoagulability) not found by PT-INR or ROTEM EXTEM can be evaluated.

Meanwhile, in the assays triggering the extrinsic coagulation pathway (EX, LI and PI), a tendency to prolongation was found in CT3 one week after surgery or one month after surgery, rather than just at the time before surgery, which matched the results of PT-INR or ROTEM EXTEM. In other words, it can be said that the risk of bleeding due to a reduced amount of FVII is reflected.

In particular, it was found that there is a risk of thrombosis and a risk of bleeding one week after surgery or one month after surgery, which cannot be identified by the existing normal tests. In normal tests (e.g. PT-INR or EXTEM), the clotting time is prolonged and therefore the risk of thrombosis is neglected. In fact, it is believed that the potential risk of thrombosis due to exposure of blood to minute amounts of tissue factor is increased.

Note that the rate of acceleration of the coagulation reaction is considered to be slow due to the decreased amount of FVII. However, there is still sufficient FVII for the coagulation reaction to proceed. Compared to the fact that the risk of a coagulation reaction in healthy blood vessels is smaller due to less tissue factor exposure in healthy people, it can be said that measurements one week after surgery and one month after surgery indicate a higher risk of thrombosis. Also, once the blood vessel is ruptured and bleeding occurs, the amount of FVII decreases and it takes time to stop bleeding. Even if hemostasis is eventually achieved, it can be fatal because no rapid clotting response occurs. During bleeding, blood is exposed to an excess of tissue factor expressed extravascularly, and therefore, the coagulation reaction proceeds rapidly in healthy people (the amount of FVII is not reduced). However, in samples corresponding to measurements one week after surgery and one month after surgery, time is required to stop bleeding even if blood is exposed to an excessive amount of tissue factor due to the decreased amount of FVII.

Furthermore, if it is possible to confirm by performing an endogenous assay IN that there are no abnormalities IN the endogenous pathway, both the thrombosis risk and the bleeding risk described above are supported by the exogenous pathway, i.e. exposure of a trace of tissue factor to the blood and reduction of the quantity of FVII by the exposure. Furthermore, they can also be distinguished from the risk of thrombosis due to a decrease in the activity of antithrombin described in example 2 described below.

Based on the above findings, focusing again on the measurement results of each coagulation factor, it can be concluded that the results do not contradict the DBCM assessment of increased risk of thrombosis due to blood exposure to trace tissue factor. In particular, although the amount of FVII decreases significantly one week after surgery and one month after surgery, other factors have little such tendency. Such a result is worth paying attention in the following point. In particular, it is a consumable possibility that a reduction in the amount of FVII cannot be neglected, since it is unlikely that only hepatic synthesis of FVII is selectively reduced compared to other coagulation factors. It is believed that very low concentrations of tissue factor are continuously exposed to the blood to the extent that no thromboembolic events occur. At such low concentrations, tissue factor does not immediately manifest as thrombosis or Disseminated Intravascular Coagulation (DIC). However, it is believed that tissue factor binds to FVII or FVIIa, which results in a consumable decrease in the amount of FVII.

One of the reasons why this risk could not be assessed by the PT-INR assay or the ROTEM EXTEM assay was thought to be due to the high concentration of tissue factor used for the assay. The final concentration of tissue factor in the EXTEM assay was estimated to be about 30 times the DBCM concentration performed this time, while the final concentration in the PT-INR assay was estimated to be about 1000 times the DBCM concentration performed this time. In these assays, an excess of tissue factor is input as a test reagent compared to the amount of tissue factor in blood. For this reason, it is believed that the hypercoagulative effects caused by tissue factor in blood are masked and, in contrast, the effect of the trend of factor FVII deficiency on the test becomes dominant, which is considered to be coagulation prolongation. Further, in ROTEM, as a characteristic of viscoelasticity measurement, soft fibrin gel, which is seen in an early stage of a coagulation reaction, is broken, and such an initial coagulation reaction cannot be detected. Therefore, it is believed that the hypercoagulability of a specimen cannot be evaluated in the case where the specimen, which is initially short in the starting time of coagulation and in a hypercoagulable state, does not rapidly turn into a hard gel due to a low level of FVII.

From the above, it is suggested that the following requirements should be put forward in order to evaluate the thrombosis risk with high accuracy.

In this device, a method (such as DBCM) that does not break the original fibrin gel without applying shear stress to the blood being measured is advantageous. In contrast, methods that disrupt the original fibrin gel during measurement (such as thromboelastometry) are disadvantageous. In addition, as the reagent, tissue factor triggering the extrinsic coagulation pathway is used at a very weak concentration. The analysis unit of the device assesses the risk of thrombosis (advantageously, the risk of thrombosis and the risk of bleeding) on the basis of the complex permittivity at a specific frequency within a predetermined time period measured at the above-mentioned time interval after the anticoagulation action on the blood sample is released. In this analysis, it is advantageous to use a feature quantity extracted from a complex permittivity spectrum of a specific frequency.

Note that the above evaluation is finally obtained by performing experiments of a large number of items when considering the present technology, and it is difficult to perform similar evaluation without the present technology. Furthermore, in normal medical practice, such exhaustive trials and trial-based assessments are difficult to perform in view of the medical economics.

< < example 2 >)

With respect to the physiological activity of antithrombin, which has been difficult to evaluate in the existing tests, studies have been made on methods for evaluating the risk of thrombosis, the risk of bleeding, and the coexistence of both risks. The changes associated with blood preservation of healthy persons were used as a validation model in experiment 1, and it was validated in experiment 2 that the addition of antithrombin to blood of healthy persons prolonged the clotting time.

< Experimental method >

The experimental procedure of experiment 1 is described below. Venous blood of healthy persons was collected by a vacuum blood collection tube using citric acid as an anticoagulant. The first tube was discarded and the blood collected in the second and third tubes was used for the experiment. Of the two blood collection tubes used for the experiment, the first was used for the experiment within one hour after blood collection. The other tube was stored at room temperature (25 ℃) for 25 hours without opening and then used for the experiment. Measurements of DBCM, PT-INR, APTT and antithrombin activity were performed. Wherein measurements other than DBCM measurements are performed using plasma obtained by centrifuging blood. Further, the assays used IN the DBCM measurement include CA assay, IN2 assay, IN4 assay, and IN20 assay. Note that the CA assay is an assay using only calcium salt as a reagent. IN addition, calcium salt and ellagic acid, which is a coagulation initiator of the intrinsic coagulation pathway, were used as reagents IN the IN2 assay, IN4 assay, and IN20 assay. Specifically, for 200 μ L of blood, 6-fold dilution of actin FSL (reagent for APTT measurement) purchased from cismexican Corporation (Sysmex Corporation) and PBS in amounts of 1.2 μ L, 2.4 μ L, and 12 μ L was used as ellagic acid.

The experimental procedure of experiment 2 is described below. Blood of a healthy person collected similarly to experiment 1 in which purchased antithrombin was added as a reagent and blood of a healthy person collected similarly to experiment 1 in which physiological saline was added as a control were prepared to perform experiments. Measurements of DBCM, APTT and antithrombin activity were performed. Among them, IN the DBCM measurement, CA measurement and IN20 measurement are performed.

< results and discussion >

The results and discussion of experiment 1 are described below. For example, the response obtained by the DBCM measurement was analyzed focusing on time CT0 (time giving the maximum value of the complex dielectric constant at 1 MHz) and time CT (time giving the minimum value of the complex dielectric constant at 10 MHz), and the results are shown in table 2 below. The results of the PT-INR, APTT and antithrombin activity measurements are also summarized in Table 2 below.

[ Table 2]

TABLE 2 blood preservation at Room temperature and test results relating to blood coagulation

According to the literature (Ann Clin biochem.2017Jul; 54(4): 448-. Other coagulation factors include coagulation factors in which the amount of FVIII is slightly reduced and coagulation factors in which the amount of FVIII is not significantly reduced. In other words, it has been generally recognized that the coagulation ability as a whole is slightly reduced because the antithrombin activity as a coagulation inhibition system is not changed and some factors of the coagulation system are reduced.

In the experimental results shown in Table 2 above, the antithrombin activity did not decrease even after blood preservation, and the time of PT-INR and APTT was prolonged by blood preservation, which is in match with the above-mentioned conventional general knowledge. However, considering the measurement results of DBCM, IN the CA assay, which does not perform artificial acceleration of the coagulation response, and IN2 and IN4 assays, IN which the intrinsic coagulation pathway is triggered very weakly, the time CT and time CT0 are significantly reduced due to blood preservation. This means that a decrease in the rate of antithrombin reaction predominates over a decrease in the coagulation response of blood preservation. Whether a very weak trigger of the coagulation reaction leads to thrombosis is a problem in terms of whether thrombosis occurs in a blood vessel in vivo. In this case, it can be said that the decrease in the reaction rate of antithrombin is a very important factor. However, such findings have not been obtained until now.

Meanwhile, it was also found that if the activation of artificial coagulation is enhanced to the extent IN the IN20 assay, no decrease IN the reaction rate of antithrombin is detected. In particular, when the triggering of the coagulation reaction is strong, the activity of the coagulation factor becomes dominant.

Furthermore, in order to quantitatively evaluate the decrease in the reaction rate of antithrombin, it is only necessary to perform measurements under various conditions that trigger coagulation and to check how much the difference in the coagulation time is according to the magnitude of the trigger. For example, it is suggested to perform the following analysis. IN the measurements, the IN20 assay or the assay that triggers the coagulation reaction more strongly than the IN20 assay and the CA assay or the assay that does not trigger the coagulation reaction strongly, such as the IN2 assay and the IN4 assay, was performed. The overall activity of the coagulation factors is then assessed from the clotting time (e.g., CT) of the strong trigger assay. If the time is longer than the reference, a decrease in the blood coagulation ability can be seen. Next, the clotting time of the weak trigger assay was evaluated. If the time is sufficiently longer than the strong trigger assay, it can be seen that there is no fear of a decrease in the antithrombin activity. In contrast, when the degree of elongation is low, the following three possibilities (and combinations thereof) may be considered as factors of the risk of thrombosis. Specifically, the factors of the risk of thrombosis correspond to one or more of a decrease in the activity of antithrombin, an endogenous hypercoagulability, and a case where the reaction of the extrinsic pathway proceeds compared with the reaction measured by an endogenous weak trigger due to the large amount of exposure of the tissue factor to blood as shown in the above example 1.

Fig. 12 is a block diagram showing an example of a coagulation analysis method in view of thorough finding of a thrombosis risk. Note that in fig. 12, evaluation criterion 1, evaluation criterion 2, and evaluation criterion 3 may be determined in advance based on the measurement results of a certain number of healthy persons and patients. Further, these references may simply be set as thresholds. More advantageously, however, the reference may be given in terms of the coagulation time(s) measured using a strong trigger assay and the coagulation time (w) measured using a weak trigger assay, respectively.

As shown in fig. 12, after the applying step and the measuring step, the detection time CT is determined for each. Specifically, for example, the endogenous weak trigger assay ct (inw), the endogenous strong trigger assay ct (ins), and the exogenous weak trigger assay ct (exw) are detected. Thereafter, determination is performed using an evaluation criterion set in advance. Specifically, for example, it is determined whether (i) assay ct (ins) is less than evaluation reference 1, (ii) a value obtained by subtracting assay ct (ins) from assay ct (inw) is less than evaluation reference 2, and (iii) assay ct (exw) is less than evaluation reference 3.

Then, using these determination results, in the case where the above (i) and (ii) are yes, a decrease in the physiological activity of antithrombin (including a decrease in the reaction rate) and/or exposure to tissue factor, etc. (risk of thrombosis) is determined, and in the case where the above (i) is yes and the above (ii) is no, it is determined that there is no abnormality in the endogenous pathway and antithrombin. In this case, if the above (iii) is yes, the exogenous hypercoagulability (thrombosis risk) due to exposure of tissue factor or the like is determined.

Further, in the case where the above-mentioned (i) is no and the above-mentioned (ii) is yes, reduction of the activity of intrinsic coagulation factor (bleeding risk) + reduction of the physiological activity of antithrombin (including reduction of the reaction rate) and/or exposure of tissue factor and the like (thrombosis risk) is determined, and in the case where the above-mentioned (i) and (ii) are no, reduction of the activity of intrinsic coagulation factor (bleeding risk) is determined. In this case, if the above (iii) is yes, it is determined that the decrease in the activity of the intrinsic coagulation factor (bleeding risk) and the extrinsic hypercoagulability (thrombosis risk) due to exposure of the tissue factor or the like coexist.

As described above, it was shown that the decrease in the rate of antithrombin reaction due to blood preservation can be assessed by the CA assay of DBCM or the assay that triggers the coagulation reaction very weakly. It is also postulated that the decrease in the rate of antithrombin reaction is due to some cause in vivo. For these reasons, it is difficult to assess the increased risk of thrombosis by existing tests or combinations thereof.

The results and discussion of experiment 2 are described below. The measurement results are summarized in table 3 below.

[ Table 3]

[ Table 3 antithrombin addition test ]

As shown in table 3 above, it was confirmed that the coagulation time was prolonged by the excessive addition of antithrombin.

EXAMPLE 3

As described above, in example 1, it was shown that even in the case of samples having both risk of thrombosis and risk of bleeding due to exposure to tissue factor and due to exposure to reduced amounts of FVII, both of these risks can be evaluated from the evaluation of exogenously determined CT and CT3, e.g. measured by DBCM. Further, in example 2, a method of evaluating the risk of thrombus formation due to reduction in antithrombin activity or the like is shown, and it has been shown that the determination thereof can be performed as shown in fig. 12. However, in fig. 12, it is difficult to completely determine whether the factors of the risk of thrombosis and/or the risk of bleeding are in the exogenous pathway, the endogenous pathway, or AT.

In this regard, assuming a plurality of cases including a case where the cause of the risk of thrombosis is due to the exposure of tissue factor (TF +) and a case where the cause of the risk of thrombosis is due to the decrease in the activity of antithrombin (AT-), how the experimental results of DBCM change is verified and summarized in table 4 below. Note that, with respect to (TF +), the item "TF + (no reduction in FVII)" is also set assuming that the reduction in the amount of FVII is not so significant.

[ Table 4]

TABLE 4 relationship between the causes of thrombosis/hemorrhage Risk and DBCM test values

In addition: low sensitivity

As shown in table 4 above, by using a variety of assays, a more detailed analysis of the risk of thrombosis and/or bleeding and their causes can be performed.

EXAMPLE 4

According to the results of example 3, a measurement (evaluation) panel assuming a more specific case was set. More specifically, a situation is assumed where there is a concern about the risk of bleeding (in perioperative periods, during anticoagulation therapy, when there is a finding suggesting bleeding, etc.).

In this case, both exogenous and endogenous assays must be performed. Either a weak trigger assay or a strong trigger assay may be used. However, in case it is desired to determine the risk of bleeding due to a reduced amount of FVII factor (due to contamination with tissue factor) or due to a reduced amount of exogenous factor in general, it is best to use a weak trigger assay as the exogenous assay. In cases where it is not necessary to perform this degree of determination and it is desirable to know the test results, a strong trigger assay may be used. For example, fig. 13 is a block diagram illustrating a coagulation analysis method for determining a bleeding risk using an exogenous weak trigger assay and an endogenous weak trigger assay.

As shown in fig. 13, after the applying step and the measuring step, the feature amount is detected for each measurement. Specifically, for example, endogenous weak trigger assays CT (in) and exogenous weak trigger assays CT (ex) and CT3(ex) are detected. Note that measuring CT (in) may be measuring CT3(in), measuring CT1(in), or a parameter based thereon. Likewise, measured CT (ex) and measured CT3(ex) may each be parameters according to this.

Thereafter, determination is performed using an evaluation criterion set in advance. Specifically, for example, it is determined whether (i) assay CT (in) is greater than evaluation reference B1, (ii) assay CT (ex) is greater than evaluation reference B2, (iii) assay CT (ex) is less than evaluation reference B3, and (iv) assay CT3(ex) is greater than evaluation reference B4.

Then, using these determination results, in the case where the above-mentioned (i) is yes, it is determined that there is a bleeding risk due to the deficiency of intrinsic coagulation factor, in the case where the above-mentioned (i) is no, it is determined that there is no bleeding risk due to the deficiency of intrinsic coagulation factor, in the case where the above-mentioned (ii) is yes, it is determined that there is a bleeding risk due to the deficiency of extrinsic coagulation factor, and in the case where the above-mentioned (ii) is no, it is determined that there is no bleeding risk due to the deficiency of extrinsic coagulation factor. Four conclusions can be reached by combining the two determinations. For example, in the case where the above (i) and (ii) are yes, it is determined that there is a bleeding risk due to the deficiency of intrinsic coagulation factors and extrinsic coagulation factors.

Further, in the case where the above-mentioned (iii) and (iv) are yes, it is determined that the risk of thrombosis due to contamination with tissue factor in blood and the risk of bleeding due to deficiency of only FVII in extrinsic coagulation factor coexist. Note that, in the case where the above (iii) is yes and the above (iv) is no, it is determined that there is a risk of thrombus formation due to extrinsic hypercoagulability.

Note that the present technology may also adopt a configuration.

(1) A coagulation system analysis device comprising:

circuitry configured to:

performing a first coagulation-related assay on a first blood sample;

performing a second coagulation-related assay on a second blood sample; and is

Assessing the activity of the coagulation factor and the activity of the coagulation inhibiting factor based at least in part on the output of the first coagulation related assay and the output of the second coagulation related assay,

wherein the first blood sample and the second blood sample are dispensed from the same blood sample collected from the patient.

(2) The coagulation system analysis device according to (1), wherein the circuit is further configured to output information indicative of a risk of thrombosis of the patient and/or information indicative of a risk of bleeding of the patient based at least in part on the output of the first coagulation-related assay and the output of the second coagulation-related assay.

(3) The coagulation system analysis device according to (1), wherein the first coagulation-related assay comprises an extrinsic coagulation pathway trigger assay, and the second coagulation-related assay comprises an intrinsic coagulation pathway trigger assay.

(4) The coagulation system analysis device according to (3), wherein the extrinsic coagulation pathway trigger assay comprises an extrinsic weak trigger assay and/or the intrinsic coagulation pathway trigger assay comprises an intrinsic weak trigger assay.

(5) The coagulation system analysis device according to (1), wherein the first coagulation-related assay and the second coagulation-related assay are selected from the group consisting of an assay that does not trigger a coagulation reaction, an endogenous strong-trigger assay, an endogenous weak-trigger assay, an exogenous strong-trigger assay, and an exogenous weak-trigger assay.

(6) The coagulation system analysis device according to (1), wherein the first coagulation-related assay comprises a strong trigger assay and the second coagulation-related assay comprises a weak trigger assay.

(7) The coagulation system analysis device according to (1), wherein the output of the first coagulation-related assay and/or the output of the second coagulation-related assay comprises a coagulation time.

(8) The coagulation system analysis device according to (1), wherein,

the first coagulation-related assay comprises an endogenous weak trigger assay configured to output a first coagulation time,

the second coagulation-related assay comprises a strong endogenous trigger assay configured to output a second coagulation time, and

the circuit is further configured to perform a third coagulation-related assay comprising an extrinsic weak trigger assay configured to output a third coagulation time.

(9) The coagulation system analysis device according to (8), wherein the determining of the activity of the coagulation factor and the activity of the coagulation inhibitory factor includes:

(i) comparing the first clotting time to a first threshold;

(ii) comparing the second clotting time to a second threshold;

(iii) comparing the third clotting time to a third threshold; and is

Outputting information indicative of a risk of thrombosis of the patient and/or information indicative of a risk of bleeding of the patient based at least in part on one or more of the comparisons (i), (ii), and (iii).

(10) The coagulation system analysis device according to (1), wherein the circuit is further configured to:

determining a first permittivity spectrum based on an output of the first coagulation-related assay;

extracting values of one or more features from the first permittivity spectrum;

determining a second permittivity spectrum based on an output of the second coagulation related assay;

extracting values of one or more features from the second permittivity spectrum; and is

Assessing the activity of the coagulation factor and the activity of the coagulation inhibitory factor based on the values of the one or more features extracted from the first permittivity spectrum and the values of the one or more features extracted from the second permittivity spectrum.

(11) The coagulation system analysis device according to (10), wherein the one or more characteristics include: the time CT at which the dielectric constant has a minimum value and the time CT3 at which the slope of the dielectric constant is maximum.

(12) The coagulation system analysis device according to (1), wherein the circuit is further configured to:

providing information associated with the output of the first coagulation related assay and information associated with the output of the second coagulation related assay as inputs to a training model, and

wherein the activity of the coagulation factor in the blood sample and the activity of the coagulation inhibitor in the blood sample are based at least in part on the output of the training model.

(13) The coagulation system analysis device according to (12), wherein,

the information associated with the output of the first coagulation related assay comprises values of one or more features extracted from a permittivity spectrum determined based on the output of the first coagulation related assay; and is

The information associated with the output of the second coagulation related assay includes values of one or more features extracted from a permittivity spectrum determined based on the output of the second coagulation related assay.

(14) The coagulation system analysis apparatus according to (12), wherein the training model comprises a model trained using one or more machine learning techniques.

(15) The coagulation system analysis device according to (1), wherein the circuit further comprises:

a measurement unit configured to perform a first coagulation-related assay and a second coagulation-related assay, the measurement unit being configured to measure a dielectric impedance of the first blood sample and the second blood sample when each blood sample is arranged between the electrodes to which the alternating voltage is applied.

(16) A method of analyzing a blood sample to determine an amount of risk of thrombosis and/or an amount of risk of bleeding in a patient associated with the blood sample, the method comprising:

performing a first coagulation-related assay on a first blood sample;

performing a second coagulation-related assay on a second blood sample; and is

Assessing the activity of the coagulation factor and the activity of the coagulation inhibiting factor based at least in part on the output of the first coagulation related assay and the output of the second coagulation related assay,

wherein the first blood sample and the second blood sample are dispensed from the same blood sample collected from the patient.

(17) A coagulation analysis system comprising:

a first electrode and a second electrode arranged opposite to the first electrode such that a container comprising a first blood sample may be arranged between the first electrode and the second electrode;

a voltage generator configured to apply an alternating voltage to the first electrode and the second electrode; and

circuitry configured to:

performing a first coagulation-related assay on a first blood sample while applying an alternating voltage to the first electrode and the second electrode;

performing a second coagulation-related assay on a second blood sample; and is

Assessing the activity of the coagulation factor and the activity of the coagulation inhibiting factor based at least in part on the output of the first coagulation related assay and the output of the second coagulation related assay,

wherein the first blood sample and the second blood sample are dispensed from the same blood sample collected from the patient.

It should be understood by those skilled in the art that various modifications, combinations, sub-combinations and alterations may be made depending on design requirements and other factors as long as they are within the scope of the appended claims or their equivalents.

List of reference marks

100 blood coagulation system analyzer

1a, 1b a pair of electrodes

101 electric measuring container

102 connection unit

103 container holding unit

2 applying unit

3 measuring cell

4 analysis unit

5 output unit

6 display unit

7 memory cell

8 measurement condition control unit

9 temperature control unit

10 blood sample supply unit

11 reagent supply unit

12 precision management unit

13 drive mechanism

14 sample Standby Unit

15 stirring mechanism

16 user interface

And 17, a server.