阅读说明：本技术 自动化液相免疫反应分析装置及其方法 (Automatic liquid phase immune reaction analysis device and method thereof ) 是由 崔義烈 朱厚暾 金亨勋 曺周铉 任旭彬 吴茔进 任伦兑 郑志雲 于 2019-07-19 设计创作，主要内容包括：本发明涉及一种适合基于酶联免疫吸附测定(ELISA)检测生物样品等中包含的特定成分的液相免疫分析装置及其方法。(The present invention relates to a liquid phase immunoassay device and a method thereof suitable for detecting specific components contained in a biological sample or the like based on enzyme-linked immunosorbent assay (ELISA).)

1. An automated liquid phase immunoassay method comprising the steps of:

dropping a washing tip into which a magnetic beam is inserted into a sample solution containing magnetic beads to trap the magnetic beads in the sample solution on a surface of the washing tip;

moving the washing tip with the magnetic beads trapped on the surface to a washing solution and putting the washing tip into the washing solution;

driving a driving motor connected to the magnetic beam to move the magnetic beam upward and move the washing tip up and down a plurality of times to disperse the magnetic beads trapped to the washing tip in the washing solution; and

the magnetic beam is inserted into a wash tip to capture magnetic beads within the wash solution.

2. The automated liquid-phase immunoassay method according to claim 1, wherein,

the step of trapping magnetic beads comprises the steps of:

in the sample solution containing the magnetic beads, a pipette arm to which the washing tip is attached and the magnetic beam are integrally moved while moving the washing tip into which the magnetic beam is inserted up and down.

3. The automated liquid-phase immunoassay method of claim 1, further comprising the steps of:

a dismounting plate is arranged below the suction pipe arm provided with the washing tip, and the dismounting plate is provided with a dismounting hole formed with a concave part;

a suction pipe arm provided with the washing tip passes through the dismounting hole;

disposing the recess of the detaching plate above the upper end of the washing tip; and

separating the wash tip from the pipette arm by moving the pipette arm to an upper portion with respect to a disassembly plate.

4. The automated liquid-phase immunoassay method of claim 1, further comprising the steps of:

moving the magnetic beads with the impurities removed to a detection chamber;

disposing an optical reader below the detection chamber; and

the optical interpreter optically inspects the sample of the detection chamber.

5. The automated liquid-phase immunoassay method of claim 4, further comprising the steps of:

arranging the standard block above the optical interpretation instrument;

the optical interpretation instrument optically inspects the fluorescence measurement standard substance in the standard block; and

comparing the result of the optical inspection of the standard substance with the result of the optical inspection of the sample of the detection chamber.

6. The automated liquid-phase immunoassay method of claim 1, further comprising the steps of:

fixing the dispensing tip at the lower part of a hollow collecting arm with a through hole in the dispensing tip;

moving a moving body to which the collection arm is fixed, so as to put the dispensing tip into a sample solution;

applying a suction force to the dispensing tip by operating a hollow pump unit connected to the collection arm to collect a sample from a sample chamber;

moving the collected sample to a reaction chamber; and

a discharge force is applied to the dispensing tip by operating the pump unit to discharge and dispense a sample into the reaction chamber.

7. The automated liquid-phase immunoassay method of claim 6, further comprising the steps of:

maintaining the sample dispensed into the reaction chamber at a constant temperature to incubate the sample.

8. The automated liquid-phase immunoassay method of claim 7, further comprising:

a first reaction step of performing a dispensing operation on a sample in a first cuvette of the plurality of cuvettes and starting incubation;

a second reaction step of performing a dispensing operation on a sample in a second cuvette of the plurality of cuvettes and starting incubation; and

a first washing step of performing a washing operation on a sample of the first cuvette.

9. The automated liquid-phase immunoassay method according to claim 8, wherein,

after the first reaction step, further comprising the steps of:

detaching a dispensing tip used in dispensing a sample in the first cuvette from the collection arm; and

mounting a dispensing tip on the collection arm to be used for dispensing a sample in the second cuvette,

after the second reaction step, further comprising the steps of:

detaching a dispensing tip used in dispensing a sample in the second cuvette from the collection arm; and

a washing tip to be used for washing the sample in the first cuvette is mounted on the pipette arm.

10. An automated liquid phase immunoassay device comprising:

a suction pipe arm, which can fix the washing tip at the lower part and has a hollow part which is communicated up and down in the suction pipe arm;

the magnetic beam is positioned in the hollow of the straw arm and can move up and down;

a moving body to which the suction pipe arm is fixed;

a moving body driving unit that moves the moving body;

a driving motor that moves the magnetic flux; and

and a control unit that controls the moving body drive unit.

11. The automated liquid-phase immunoassay device according to claim 10,

and the lower part of the magnetic beam is provided with a permanent magnet.

12. The automated liquid phase immunoassay device of claim 10, further comprising:

a collection arm to which a dispensing tip is fixed at the lower part thereof, which has a hollow part penetrating vertically therein, and which is fixed to the movable body; and

a pump unit connected to the hollow of the collection arm and capable of providing a suction force or a discharge force to the dispensing tip.

13. The automated liquid-phase immunoassay apparatus according to claim 10, further comprising a punch arm provided with a punch tip at a lower portion and fixed to the moving body,

the length from the moving body to the lower portion of the punching arm is longer than the length from the moving body to the lower portion of the straw arm.

14. The automated liquid-phase immunoassay device of claim 10, further comprising a detachment plate provided with a detachment hole forming a recess,

the area of the disassembly hole is larger than that of the upper end of the washing tip.

15. The automated liquid phase immunoassay device of claim 10, further comprising:

a holder having a groove-shaped mounting channel to which one or more cuvettes can be mounted and an inspection hole that penetrates in the vertical direction; and

and a holder driving part which can adjust the position of the holder.

16. The automated liquid-phase immunoassay device according to claim 15,

the holder comprises a hot plate in the lower part for maintaining the cuvette at a constant temperature.

17. The automated liquid phase immunoassay device of claim 15, further comprising:

the optical interpretation instrument is provided with a light source, a beam splitter, a lens and a detector; and

an interpreter driving section that moves a position of the optical interpreter to align with the inspection hole of the holder.

18. The automated liquid-phase immunoassay device according to claim 15,

the holder includes:

and a standard block which is provided with an optical hole penetrating in the vertical direction and can be loaded with a fluorescence measurement standard substance.

Technical Field

The present invention relates to an analysis system or apparatus for detecting a specific component contained in a biological sample by Enzyme Linked immunosorbent Assay (ELISA) liquid-phase immunoassay, and a method thereof.

Background

With the development of the medical and bioengineering fields and various related technologies, examinations for detecting various molecular indicators such as blood cells, genes, proteins, antigens, pathogens, etc. in predetermined biological samples such as urine and blood have been widely performed. Typically, the inspection process includes: after the sample is collected, the collected sample is reacted with a predetermined reagent suitable for the target index, and then the change is analyzed and observed. Thereby, qualitative and/or quantitative analysis of various molecular indicators contained in the sample can be performed, and based thereon information on the diagnosis, progression status or prognosis of the disease can be obtained.

One of the techniques widely used in such examination procedures is an immunoreaction technique based on specific binding between antigen/antibody, also known as Enzyme Immunoassay (EIA). Among them, depending on the kind of the substrate used for detecting the analyte, there are a color change measurement method (chromogenic method or colorimetric method) in which a color reaction is measured by absorbance, a chemiluminescence method, a method using fluorescence, and the like. In addition, a sandwich type immune response or a competitive type immune response called Enzyme Linked Immunosorbent Assay (Enzyme Linked Immunosorbent Assay) is included according to the Assay format.

In such an analysis, regardless of the mode used, it is preferable to remove a non-specific reactant for highly specific and highly sensitive detection. That is, in the examination, after a reaction between a reagent and a sample occurs, in order to accurately detect a reaction product, it is necessary to purify or separate (purify) the reaction product. However, in many cases, in order to detect the reaction product, it is also necessary to use a membrane such as nitrocellulose or to use a two-dimensional flat plate. However, the use of such a membrane or plate not only limits the reaction area, but also makes it difficult to remove non-specific products.

The most effective method for removing non-specific reactants is physical washing or purification. Therefore, it is required to develop a device/system capable of accurately and rapidly performing a plurality of examinations for quantifying the reaction of a sample and a reagent, physical purification, detection, and interpretation/analysis of a reaction product in one integrated system.

Documents of the prior art

Patent document

Patent document 1: korean laid-open patent No. 10-2012 and 0027359 (published: 03 month and 21 days 2012)

Patent document 2: korean laid-open patent No. 10-2016-

Patent document 3: korean laid-open patent No. 10-2018-0079150 (published: 2018 year 07 month 10 day)

Disclosure of Invention

Technical problem to be solved

It is an object of the present invention to provide a device or system and method suitable for performing a reaction of a sample and a reagent, purification/separation of a reaction product, and detection/interpretation/analysis of a reaction product comprehensively for a plurality of samples using a liquid phase-based enzyme-linked immunoassay.

The object of the present invention is not limited to the above-mentioned object, and other objects not mentioned can be clearly understood by those skilled in the art from the following description.

(II) technical scheme

One aspect of the present invention for achieving the above objects provides an automated liquid-phase immune response analysis method comprising the steps of: dropping a washing tip into which a magnetic beam is inserted into a sample solution containing magnetic beads to trap the magnetic beads in the sample solution on a surface of the washing tip; moving the washing tip with the magnetic beads trapped on the surface to a washing solution and putting the washing tip into the washing solution; driving a driving motor connected to the magnetic beam to move the magnetic beam upward and move the washing tip up and down a plurality of times to disperse the magnetic beads trapped to the washing tip in the washing solution; and inserting the magnetic beam into a wash tip to capture magnetic beads within the wash solution.

Preferably, the step of trapping magnetic beads comprises the steps of: in the sample solution containing the magnetic beads, a pipette arm to which the washing tip is attached and the magnetic beam are integrally moved while moving the washing tip into which the magnetic beam is inserted up and down.

Preferably, the automated liquid-phase immune reaction analysis method further comprises the steps of: a dismounting plate is arranged below the suction pipe arm provided with the washing tip, and the dismounting plate is provided with a dismounting hole forming a concave part; a suction pipe arm provided with the washing tip passes through the dismounting hole; disposing the recess of the detaching plate above the upper end of the washing tip; and separating the wash tip from the pipette arm by moving the pipette arm to an upper portion with respect to the detaching plate.

Preferably, the automated liquid-phase immune reaction analysis method further comprises the steps of: moving the magnetic beads with the impurities removed to a detection chamber; disposing an optical reader below the detection chamber; and the optical interpreter optically inspects the sample in the detection chamber.

Preferably, the automated liquid-phase immune reaction analysis method further comprises the steps of: arranging the standard block above the optical interpretation instrument; the optical interpretation instrument optically inspects the fluorescence measurement standard substance in the standard block; and comparing the result of the optical inspection of the standard substance with the result of the optical inspection of the sample of the detection chamber.

Preferably, the automated liquid-phase immune reaction analysis method further comprises the steps of: fixing the dispensing tip at the lower part of a hollow collecting arm with a through hole in the dispensing tip; moving a moving body to which the collection arm is fixed, so as to put the dispensing tip into a sample solution; applying a suction force to the dispensing tip by operating a hollow pump unit connected to the collection arm to collect a sample from a sample chamber; moving the collected sample to a reaction chamber; and applying a discharge force to the dispensing tip by operating the pump unit to discharge and dispense the sample to the reaction chamber.

Preferably, the automated liquid-phase immune reaction analysis method further comprises the steps of: the sample dispensed in the reaction chamber is maintained at a constant temperature to incubate (incubation) the sample.

Preferably, the automated liquid-phase immune response analysis method further comprises: a first reaction step of performing a dispensing operation on a sample in a first cuvette of the plurality of cuvettes and starting incubation; a second reaction step of performing a dispensing operation on a sample in a second cuvette of the plurality of cuvettes and starting incubation; and a first washing step of performing a washing operation on a sample of the first cuvette.

Preferably, after the first reaction step, further comprising the steps of: detaching a dispensing tip used in dispensing a sample in the first cuvette from the collection arm; and mounting on the acquisition arm a dispensing tip to be used for dispensing the sample in the second cuvette, further comprising, after the second reaction step, the steps of: detaching a dispensing tip used in dispensing a sample in the second cuvette from the collection arm; and mounting a washing tip to be used for washing the sample in the first cuvette on the pipette arm.

In addition, another aspect of the present invention provides an automated liquid-phase immunoassay device comprising: a suction pipe arm, which can fix the washing tip at the lower part and has a hollow part which is communicated up and down in the suction pipe arm; the magnetic beam is positioned in the hollow of the straw arm and can move up and down; a moving body to which the suction pipe arm is fixed; a moving body driving unit that moves the moving body; a driving motor that moves the magnetic flux; and a control unit that controls the moving body driving unit.

Preferably, the lower portion of the magnetic flux is provided with a permanent magnet.

Preferably, the automated liquid-phase immunoassay device further comprises: a collection arm to which a dispensing tip is fixed at the lower part thereof, which has a hollow part penetrating vertically therein, and which is fixed to the movable body; and a pump unit connected to the hollow of the collection arm and capable of providing a suction force or a discharge force to the dispensing tip.

Preferably, the apparatus further comprises a punch arm provided at a lower portion thereof with a punch tip and fixed to the moving body, and a length from the moving body to a lower portion of the punch arm is longer than a length from the moving body to a lower portion of the pipette arm.

Preferably, the device further comprises a disassembly plate provided with a disassembly hole formed with a depression, the disassembly hole having an area greater than that of the upper end of the washing tip.

Preferably, the automated liquid-phase immunoassay device further comprises: a holder having a groove-shaped mounting channel to which one or more cuvettes can be mounted and an inspection hole that penetrates in the vertical direction; and a holder driving part which can adjust the position of the holder.

Preferably, the holder comprises a hot plate at the lower part for keeping the cuvette at a constant temperature.

Preferably, the automated liquid-phase immunoassay device further comprises: the optical interpretation instrument is provided with a light source, a detector, a beam splitter, a lens and a detector; and an interpreter driving section that moves a position of the optical interpreter to align with the inspection hole of the holder.

Preferably, the holder includes: and a standard block which is provided with an optical hole penetrating in the vertical direction and can be loaded with a fluorescence measurement standard substance.

(III) advantageous effects

According to the automated liquid-phase immunofluorescence analysis apparatus of the present invention, the dispensing and reaction of the sample and the reagent and the separation (purification) of the reaction product by the washing module using Magnetic Beads (Magnetic Beads) are comprehensively performed, and the reaction product can be detected/interpreted with high sensitivity and high specificity using the liquid-phase sample optical system, as compared with the existing method.

In particular, according to the present invention, after a reaction of a dispensed sample, a reagent and a sample, an examination for detecting and interpreting/analyzing a reaction product is accurately and rapidly performed in one integrated system, so that it is possible to shorten an examination time, improve the accuracy and reproducibility of the examination, and reduce steps and input costs included in the entire examination.

In addition, the automated liquid-phase immunoassay device according to the present invention has a holder with a plurality of mounting channels, so that a plurality of cuvettes are coupled to one holder, and a plurality of diagnoses and analyses can be simultaneously performed within one system. Accordingly, various examinations and diagnoses/analyses can be rapidly performed to make an accurate diagnosis at a place for examination and treatment, thereby saving time, cost, and labor.

The housing included in the automated liquid-phase immunoassay apparatus according to the present invention blocks the inflow of foreign substances, so that more accurate sample inspection can be performed. And, while providing a driving unit providing up-and-down and horizontal moving forces, an optical discriminator is provided on a left-and-right moving path of the cuvette, so that sample examination can be performed through a quick and simple operation.

In addition, the pump unit included in the automated liquid-phase immunoassay device according to the present invention can accurately control the amount of a sample, a reagent, or a reaction product sucked or discharged through the dispensing tip.

In addition, unlike the gear type, the belt type front and rear driving unit included in the automated liquid phase immunoassay apparatus according to the present invention can prevent vibration and foreign substances caused by friction generated when moving left and right, thereby enabling more accurate inspection.

In addition, in the arm unit provided in the automated liquid-phase immunofluorescence analysis apparatus according to the present invention, the punch arm, the collection arm, and the pipette arm are integrally provided and formed as an integrated module, so that the up-down direction position can be controlled by one driving motor when a dispensing Pump (Pump dispenser), a punch is driven, and the dispenser tip and the washing tip are washed and separated. Therefore, unlike when each module is constituted by a respective drive motor, respectively, the size can be reduced and the manufacturing cost can be reduced.

Also, in the apparatus according to the present invention, when a plurality of cuvettes are used, a set of dispensing tip and washing tip can be used without replacing the tip for each cuvette in the middle of reaction, and the tip can be easily detached by detaching the module.

In addition, the apparatus according to the present invention includes a standard block, so that a deviation of signal values between apparatuses can be reduced.

Drawings

Fig. 1 is a photograph of the appearance of a device actually manufactured according to one embodiment of the present invention.

Fig. 2 is a schematic view showing a process of sandwich immunoreaction using magnetic beads used in a device according to an embodiment of the present invention.

Fig. 3 is a schematic diagram illustrating a competition immunoreaction process using magnetic beads used in a device according to an embodiment of the present invention.

Fig. 4 is a diagram showing the structure of a cuvette used in an apparatus according to an embodiment of the present invention.

Fig. 5 is a diagram showing a dispensing tip and a wash tip, respectively, for use in conjunction with a cuvette for use in an apparatus according to one embodiment of the invention.

Fig. 6 is a diagram showing one shape of a cuvette mounted with a dispensing tip and a washing tip used in the apparatus according to one embodiment of the present invention.

Fig. 7 is a diagram showing an appearance of an apparatus according to an embodiment of the present invention.

Fig. 8 and 9 are diagrams of the apparatus according to an embodiment of the present invention with the housing omitted.

Fig. 10 and 11 are diagrams showing a holder of the apparatus according to an embodiment of the present invention and a shape in which a cuvette is loaded in the holder.

Fig. 12 is a diagram illustrating the structure of a detachment module of an apparatus according to an embodiment of the present invention.

Fig. 13 is a diagram illustrating the rear of the interior of the device according to one embodiment of the present invention.

Fig. 14a and 14b are diagrams illustrating the structure of a dispenser (dispenser) module in an automated liquid-phase immunoassay device according to an embodiment of the present invention.

Fig. 15a is a configuration diagram showing a schematic structure of a dispenser module in an automated liquid-phase immunoassay apparatus according to another embodiment of the present invention.

Fig. 15b is an enlarged view illustrating in detail a washing tip portion of a dispenser module in an automated liquid-phase immunoassay device according to another embodiment of the present invention.

Fig. 16a and 16b are diagrams of a fluorescence optical system and a chemiluminescence optical system, respectively, that can be employed in an apparatus according to an embodiment of the invention.

Fig. 17 is a flowchart illustrating an overall flow of an automated liquid-phase immunoassay method according to an embodiment of the present invention.

Fig. 18 is a flowchart illustrating a washing process of an automated liquid-phase immunoassay method according to an embodiment of the present invention.

Fig. 19 is a flowchart illustrating an optical inspection process using a standard block in an automated liquid-phase immunoassay method according to an embodiment of the present invention.

Fig. 20 is a flowchart illustrating in detail a sample dispensing method of an automated liquid phase immunoassay method according to an embodiment of the present invention.

Fig. 21 is a timing diagram illustrating the operation of each cuvette when three cuvettes are included in the automated liquid phase immunoassay method according to one embodiment of the present invention.

Fig. 22 is a photograph showing the effect of a permanent magnet on a sample containing magnetic beads used in an automated liquid-phase immunoassay method according to an embodiment of the present invention.

Fig. 23 is a graph showing the magnetic field strength of a permanent magnet used in an automated liquid phase immunoassay method according to an embodiment of the present invention.

Fig. 24 is a photograph showing the position of a washing tip used in an automated liquid-phase immunoassay method according to an embodiment of the present invention.

Detailed Description

Hereinafter, preferred embodiments of the present invention will be described with reference to the accompanying drawings. The present embodiments are exemplary, and do not limit the invention in any way.

As shown, spatially relative terms such as "lower," "rear," "upper," and the like may be used to readily describe one element or component's relative relationship to another element or component. Spatially relative terms should be understood to include the orientation shown in the drawings and the different orientations of the elements in use or operation. For example, when an element shown in the drawings is turned over, elements described as "below" or "beneath" another element may be oriented "above" the other element. Thus, the exemplary term "below" can include both an orientation of above and below. The components may be oriented in other directions and the spatially relative terms may be interpreted based on the orientation. For example, "left-right direction" may also be interpreted as "up-down direction", and is not limited thereto.

In this specification, spatially relative terms refer to orientations when viewing the front of a device according to the invention.

In the present specification, angles and directions mentioned in describing the structure of the present invention are based on the angles and directions described in the drawings. In the description, when reference points and positional relationships with respect to angles are not explicitly mentioned in the description of structures constituting the present invention, reference is made to the relevant drawings.

Hereinafter, first, terms used in the present specification and the principle of chemical reaction used with the present apparatus will be explained.

In the present specification, "detection" refers to quantitative or qualitative analysis of a reaction product or an analyte contained after purification of the reaction product thereof after a reagent reacts with a sample to determine the presence or amount of the analyte contained in the sample described later. The detection result is interpreted by the automated liquid-phase immunoassay device 1 according to one embodiment of the present invention.

In this specification, "examination" is used as a term covering all detection, analysis and interpretation.

The term "sample" as used in this specification refers to a composition intended to contain an analyte, and a sample that can be used in the present invention is a liquid phase or a substance having fluidity similar to that of a liquid. The sample used in one embodiment of the present invention may be a biological sample, and may be, for example, a body component derived from an organism such as whole blood, plasma, serum, urine, saliva, feces, and a cell extract.

The term "analyte" as used in this specification is a compound of interest, also referred to as a target or marker, in a sample, including, but not limited to, a protein component such as an antigen or a nucleic acid substance such as a gene.

In the present specification, a "reagent" is a substance used in mixture with a sample to quantitatively or qualitatively analyze an analyte contained in the sample, and may be different depending on the kind of a specific analyte, and for example, may include, but is not limited to, a reaction buffer or buffer, a dilution buffer, a detection buffer, a washing buffer, or a predetermined antibody, enzyme, or substrate that reacts with various substances in the sample, for example, with an antigen or the like.

Fig. 1 shows the appearance of a device 1 manufactured according to one embodiment of the present invention.

The automated liquid-phase immunoassay device 1 according to one embodiment of the present invention is suitable for an immunoassay (enzyme-linked immunosorbent assay (ELISA)) based reaction based on specific binding between antigen/antibody, for example, a specific component or analyte contained in a biological sample or the like can be detected by the reaction as shown in fig. 2 and 3, and is a device suitable for physical washing, i.e., separation of unreacted materials from reaction products using magnetic beads before detecting the analyte.

Fig. 2 and 3 show ELISA analysis processes of various ways of analyzing analytes. Sandwich-type immunoreaction (Sandwich immuno assay) refers to an immunoreaction that binds a capture antibody to a detection antibody in Sandwich format by chemically binding an enzyme to the detection antibody to induce a quantitative reaction with a substrate. At this time, the capture antibody is chemically or physically bound to the magnetic beads, and the detection antibody utilizes a conjugate bound to an enzyme. The sandwich reaction using such magnetic beads can be roughly classified into two types, which are 1-step reaction (1step assay) or 2-step reaction (2step assay) depending on the number of washing steps. The method of reacting the sample with the capture antibody and then with the detection antibody after washing is referred to as a 2-step reaction, and the method of reacting with both the capture antibody and the detection antibody without distinction is referred to as a 1-step reaction (fig. 2).

In addition to sandwich-type immune responses, competitive-type responses (Competition assay), which are widely used to detect small amounts of protein molecules, are also classified into two methods. The indirect-type competition reaction or the direct-type competition reaction is classified according to whether a competitive protein or an antibody is conjugated to the magnetic beads, and the 1-step reaction and the 2-step reaction are classified according to the step of the immunoreaction. For example, fig. 3 shows one form of indirect and direct competition reactions among competition reactions.

In one embodiment according to the present invention, a fluorescent signal is used to detect the reaction product. In this case, for example, enzyme-substrate reactions such as Alkaline phosphatase (ALP) and 4-Methylumbelliferyl phosphate (MUP) are used. ALP is one of enzymes, and is a representative enzyme that causes dephosphorylation reaction. The 4-MUP reacts with ALP, dephosphorylation is irreversibly carried out by enzymatic hydrolysis, and the produced 4-Methylumbelliferone (4-MU) has a fluorescence property of being excited at a wavelength of 360nm and emitting a wavelength of 450nm, and the concentration of an analyte in a sample is determined by detecting the intensity of this fluorescence signal.

In another embodiment according to the present invention, the reaction product is detected using colorimetric methods. Colorimetric analysis is the detection of a change in the visible color of the reaction product that absorbs light at a particular visible wavelength, wherein the signal from the reaction product is used to detect absorbance to determine the concentration of the analyte in the sample. For example, typical examples of the enzyme and the substrate include peroxidase and its substrate 3,3',5,5' -tetramethylbenzidine (3,3',5,5' -tetramethylbenzidine, TMB), 3',4,4' -diaminobenzidine (3,3',4,4' -diaminobenzidine, DAB), 4-chloro-1-naphthol (4-chloro-1-naphthol, 4CN), 2 '-diaza bis [ 3-ethyl-benzothiazoline ] sulfonate (2,2' -azinodi [ 3-ethyl-benzothiazoline ] sulfate, ABTS) and o-phenylenediamine (o-phenylenediamine, OPD), but are not limited thereto. For example, when TMB is used as the substrate, blue color is generated, which can be detected by light having a wavelength of 650nm, and when ABTS is used, blue-green color is generated, which can be detected by light having a wavelength of 405nm to 410 nm. Examples of other enzyme substrates include, but are not limited to, ALP and its substrate 5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium (5-bromo-4-chloro-3-indolylphosphate/nitro blue tetrazolium, BCIP/NBT) and p-nitro-phenylphosphate (p-nitro-phenylphosphate, p-NPP). This may produce a deep yellow color and may be detected by light having a wavelength of 405nm to 410 nm.

In yet another embodiment of the present invention, chemiluminescence (chemiluminescecent) is used to detect the reaction product. Chemiluminescence is light emitted when excited electrons generated by a chemical reaction return to the ground state. Chemiluminescence does not require a light source and is measured in Relative Light Units (RLUs) per hour for determining the concentration of an analyte in a sample. Examples of the enzyme and the substrate include, but are not limited to, peroxidase and luminol as a substrate thereof, polyphenol (including pyrogallol, erythrophenol, gallic acid, umbelliferone, and the like), acridine ester (acridine ester) or fluorescein (referred to as bioluminescence when used). Other examples of the enzyme and substrate include ALP and 3- (2 '-spiroadamantane) -4-methoxy-4- (3' -phosphonooxy) -phenyl-1,2-dioxetane (3- (2'-spiroadamantyl) -4-methoxy-4- (3' -phosphonooxy) -phenyl-1,2-dioxetane, AMPPD), but are not limited thereto.

In such assays, in particular, highly specific and highly sensitive detection is required, for which removal of non-specific or unreacted substances is required. That is, in the examination process, after the reagent and the sample react, it is necessary to purify or separate the reaction product in order to accurately detect the reaction product, and the apparatus according to the present invention is an apparatus suitable for effectively removing such unreacted materials.

Specifically, the device according to one embodiment of the present invention is adapted to remove unreacted materials by using magnetic physical washing, then separate only products of specific reactions in the form of magnetic beads using a permanent magnet and concentrate, and selectively bind a detector (detector) to which an enzyme is attached to the reaction products, and finally react the enzyme with the substrates to detect signals of the reaction products thus produced.

The reaction as described above used in the device according to one embodiment of the present invention is performed in a liquid state in a cuvette mounted on the device. The apparatus according to one embodiment of the present invention is adapted to perform optimized reaction steps taking into account the characteristics of the various parameters performed in the reaction, to perform the reaction as described above in the cuvette and to detect the reaction product.

First, a cuvette 10 used in an automated liquid-phase immunoassay apparatus 1 according to an embodiment of the present invention will be described.

Fig. 4 is a diagram showing the structure of a cuvette 10 used in an automated liquid-phase immunoassay apparatus according to an embodiment of the present invention, fig. 5 is a diagram showing a dispensing tip 20 and a washing tip 30 used in the automated liquid-phase immunoassay apparatus according to an embodiment of the present invention, respectively, and fig. 6 is a diagram showing a state in which the dispensing tip 20 and the washing tip 30 are mounted on the cuvette 10 used in the automated liquid-phase immunoassay apparatus according to an embodiment of the present invention.

The cuvette 10 used in the automated liquid-phase immunoassay device 1 according to one embodiment of the present invention is used for a reaction for detecting an analyte contained in a sample, and in the cuvette, the sample reacts with a reagent to generate a reaction product, and the reaction product is washed.

As shown in fig. 4 and 6, the cuvette 10 used in the automated liquid-phase immunoassay device 1 according to one embodiment of the present invention may have an elongated shape extending in the front-rear direction. In addition, the cuvette 10 may include more than one insertion hole and a plurality of chambers. The chamber may also be referred to as a well.

Before the start of the examination or during the examination, the insertion holes are places where the wash tip 30 and the dispensing tip 20 shown in fig. 5 are inserted and wait, and are provided with the wash tip insertion hole 21 and the dispensing tip insertion hole 31, respectively.

The chambers may be configured to sequentially include a sample filling chamber 12, buffer and diluting chambers 13a, 13b, 13c, and 13d, a reaction chamber 14, a washing chamber 15, and a detection chamber 16.

Alternatively, as shown in fig. 4 and 6, the chamber may be configured to sequentially include a sample filling chamber 12, a washing tip insertion hole 21, a dispensing tip insertion hole 31, buffer and diluting chambers 13a, 13b, 13c, and 13d, a reaction chamber 14, a washing chamber 15, and a detection chamber 16.

In addition, the chamber may be sealed by a predetermined sealing film (not shown) to prevent the reagent from being denatured or contaminated, etc.

The sample filling chamber 12 is, for example, provided to be filled with various samples, such as biological samples to be analyzed, and as described above, the sample filling chamber 12 may be formed in front of or behind the wash tip insertion hole 21 and the dispensing tip insertion hole 31.

The buffer solution (also referred to as a buffer) and the chambers for dilution 13a, 13b, 13c, and 13d are filled with Magnetic Bead (MB) buffer (in the chamber 13 a), detection buffer (in the chamber 13 b), and sample dilution buffer (in the chamber 13 c) necessary for the reaction, and are disposed behind the sample filling chamber 12 or the wash tip insertion hole 21 and the dispensing tip insertion hole 31 in the above-described order to dilute the sample (in the chamber 13 d).

The reaction chamber 14 is provided to perform a reaction between a sample and a reagent, and the reaction chamber 14 is formed behind the buffer and diluting chambers.

The washing chamber 15 is a chamber that washes reaction products after the reaction in the reaction chamber, and may include a plurality of washing chambers, and in one embodiment of the present invention, three washing chambers 15a, 15b, and 15 c.

The detection chamber 16 is where the test sample reacts with the reagent to produce a reaction product, and is configured to detect the presence of the analyte in the reaction product after washing in the washing chamber 15. The detection chamber 16 is formed at the rear of the washing chamber 15, and may be provided with light transmittance for detecting a fluorescent signal.

In one embodiment of the present invention, the cuvette 10 may further include a barcode or QR code (not shown) used in conjunction with a chip described later inserted into the automated liquid-phase immunoassay device 1 of the present invention. In the present invention, barcodes include, but are not limited to, UPC-A, UPC-E, EAN, Code 39 (Code 3of 9), Interleaved 2/5 (Interleaved 2of 5), Code 128 (Code 128), UCC/EAN-128, Codabar (Codabar), postal digital Code technology barcode (PostNet), pharmaceutical Code (Pharmacode), or PDF-417, and include, but are not limited to, 1D barcodes or 2D barcodes. The barcode or QR code encodes an analyte type or the like according to a sample type.

The cuvette 10 used in the automated liquid-phase immunoassay apparatus 1 according to one embodiment of the present invention is mounted with a dispensing tip 20 and a washing tip 30.

The dispensing tip 20 may comprise a disposable microtip (e.g., a micropipette tip having a capacity of 2-1000 μ l) that is used in conjunction with a collection arm 556 described below to dispense or dispense reagents between the sample and/or the chambers described above, i.e., from one chamber to another. The dispensing tip 20 has a tubular shape, and its diameter is gradually reduced toward its tip, so that its tip portion may have a tapered shape.

The dispensing tip 20 as described above can be used with an apparatus that does not have a separate reagent supply device and a device for washing contaminants, thereby simplifying the operation of the apparatus.

The plurality of cuvettes used in the apparatus according to an embodiment of the present invention are configured such that each cuvette can be separately mounted with a dispensing tip and a washing tip, and thus can be used separately from the tips for the other cuvettes, so that contamination can be prevented. In the case of the existing automated apparatus using a metal injection needle, it is necessary to provide a means for washing the apparatus to prevent contamination, and thus, there is a problem in that the volume is increased due to the provision of an additional means, and a separate process for washing is required, thereby increasing inspection costs.

In particular, the dispensing tip 20 is inserted and mounted into the dispensing tip insertion hole 21 of the cuvette 10, and at the start of an inspection process, the dispensing tip 20 is fastened to a later-described collection arm 556 and functions as a suction or discharge together with the pump unit 506 in order to dispense or dispense a sample or a reagent between chambers. In addition, in the inspection process, in order to perform the reaction in the second cuvette or the third cuvette during the reaction in the first cuvette, the dispensing tip used in the first cuvette may be temporarily stored in the insertion hole 21, and thus, only one tip may be used in one cuvette until the inspection is completed without replacing the tip halfway, thereby having advantages of convenience and reduction of reaction time. This is explained in more detail during the operation of the device according to one embodiment of the invention.

The washing tip 30 is a member having a predetermined height and a predetermined width and having a tube shape with a lower end sealed, and an upper portion thereof is formed with an input hole having a predetermined depth and an inner diameter. The washing tip 30 is made of a non-magnetic material to be able to transfer magnetism, and may be made of a flexible material to be easily fixed to and separated from the washing arm. The wash tip 30 is also inserted and fitted into the wash tip insertion hole 21 of the cuvette 10, and at the start of the inspection process, the wash tip 30 is fastened to the pipette arm 554 to perform washing as described later. In addition, in the inspection process, in order to perform the reaction in the second cuvette or the third cuvette during the reaction in the first cuvette, the washing tip used in the first cuvette may be kept in the insertion hole 31, and thus, only one tip may be used in one cuvette, thereby having advantages of convenience and reduction in reaction time. This is explained in more detail during the operation of the device according to one embodiment of the invention.

Three cuvettes are used in one embodiment according to the invention, which are suitable for performing three types of analysis. For example, three types of different analytes in the same biological sample are analyzed, and for example, Free Thyroxine (FT 4), Thyroid Stimulating Hormone (TSH) and triiodothyronine (T3) for diagnosing Thyroid gland, and chorionic gonadotropin (hCG), Estriol (E3) and Alpha Fetoprotein (AFP) for examining malformations can be cited.

Hereinafter, an automated liquid-phase immunoassay device 1 according to an embodiment of the present invention is explained.

Fig. 7 is a diagram showing the automated liquid-phase immunoassay device 1 according to one embodiment of the present invention, and fig. 8 and 9 are diagrams showing the housing 100 omitted from the automated liquid-phase immunoassay device 1 according to one embodiment of the present invention from different directions.

The automated liquid phase immunoassay device 1 according to one embodiment of the present invention is an automated liquid phase immunoassay device 1 for inspecting a sample by inserting a cuvette 10, which may include a housing 100, a frame 200, a cuvette module 300, an optical interpretation module 400, and a dispenser module 500.

The housing 100 forms the entire exterior of the automated liquid phase immunoassay device 1 and simultaneously functions to block the flow of foreign substances into the interior thereof.

The case 100 may be provided with various input portions for operation and a display portion 110 for output. In addition, the case 100 is provided with an access port 120 into which the cuvette 10 is inserted. When the cuvette 10 enters the inside of the housing 100 through the access port 120, the flow of foreign substances into the chamber included in the cuvette 10 is blocked by the housing 100, and thus more accurate sample inspection can be performed.

A frame 200 may be provided within the housing 100 to fix the cuvette module 300, the optical interpretation module 400, the dispenser module 500, and the like. The frame 200 may include a lower frame 210, a first side frame 220, a second side frame 230, and a rear frame 240.

The lower frame 210 is disposed at the lower portion of the automated liquid-phase immunoassay device 1. The lower frame 210 may have a plate-shaped structure having a predetermined area.

The first and second side frames 220 and 230 are respectively disposed at left and right sides of the lower frame 210, and may be erected at a predetermined height. In addition, the first and second side frames 220 and 230 may have guide spaces 222, 232 that guide the displacement of the holder 310 in the front-rear direction, respectively.

The rear frame 240 is located at the rear of the apparatus, and may be provided so that a predetermined control device or the like can be fixed.

Fig. 10a, 10b and 11 are diagrams showing a holder 310 of an automated liquid-phase immunoassay device according to an embodiment of the present invention and a shape in which cuvettes 10 are loaded in the holder 310. Fig. 12 is a diagram showing the structure of a detachment module 340 in an automated liquid-phase immunoassay device according to an embodiment of the present invention. Fig. 13 is a diagram showing the rear of the interior of an automated liquid-phase immunoassay device according to an embodiment of the present invention.

The cuvette module 300 will be explained below.

The cuvette module 300 is disposed within the housing 100, and the cuvette module 300 is a device that receives the cuvettes 10 and moves the received cuvettes 10 in the front-rear direction.

The cuvette module 300 may include a holder 310, a holder driving part 320, a holder guide 330, and a detachment module 340.

The holder 310 is a component to which the cuvette 10 can be mounted. For example, the holder 310 may be provided on the lower frame 210 and may be provided behind the access 120 of the housing 100. Thus, the cuvette 10 may be pushed into the holder 310 through the access opening 120.

On the other hand, the holder 310 may have a groove-shaped mounting channel 312 to enable insertion and mounting of more than one cuvette 10, respectively. The mounting channel 312 may extend in the front-rear direction and open to the front.

An inspection hole 314 is formed at the rear end of the mounting channel 312. The inspection hole 314 is a portion formed to penetrate in the vertical direction. Therefore, when the cuvette 10 is received and mounted in the mounting passage 312 of the holder 310, a lower portion of a rear portion of the holder 310 is exposed downward through the inspection hole 314. Specifically, the lower portion of the detection chamber 16 disposed behind the cuvette 10 may be exposed downward through the inspection hole 314.

In addition, a plurality of the mounting channels 312 are formed in the holder 310, so that cuvettes 10 can be inserted into each of the mounting channels 312 and a plurality of cuvettes 10 can be inspected. At this time, a plurality of the installation channels 312 may be laterally arranged side by side with each other in one holder 310.

A hot plate 316 and a hot plate power supply unit 318 are provided at the lower portion of the holder 310. This is to automatically control the cuvette and the reactants in the cuvette to be maintained at a constant temperature during the reaction, and to ensure the accuracy and precision of the examination according to the characteristics of the biological sample sensitive to temperature.

The hot plate 316 heats the holder 310 to heat the cuvette 10 and the sample and the reactants contained therein to a constant temperature and maintain a specific temperature by convection. The temperature is automatically controlled by a built-in program. Temperature sensors are employed for automatic control and in one embodiment of the invention, temperature sensors are used inside the holder, hot plate and device. Since the temperature inside the device affects the optical system, the temperature sensor of the device is used to control the temperature inside the device. The temperature sensor of the hot plate controls the temperature of the hot plate, and the temperature sensor of the holder measures the temperature of the holder and controls the hot plate in a feedback manner.

The holder driving part 320 may adjust the position of the holder. In one embodiment of the present invention, the holder driving part 320 may be composed of a member that applies force to the holder 310 in the front and rear direction. The holder driving part 320 may include a movable body 322 to which the holder 310 is fixed, a driving motor, and a predetermined transmission member that transmits power of the driving motor to the movable body 322. The driving motor may use a servo motor, a stepping motor, a Direct Current (DC) motor, or the like.

The holder guide 330 is provided to guide the front-rear direction displacement of the holder 310. The holder guide part 330 may include: a predetermined guide rail extending in the front-rear direction; and a predetermined guide portion connected to the guide rail, and movable forward and backward along the guide rail and connected to the movable body 322.

The detachment module 340 is a component for dispensing/mixing reagents in other cuvettes during an immunoreaction time (incubation) after using a dispensing tip and washing the tip in an immunoassay, or for detaching the tip after the reaction in each cuvette is finished.

The detachment module 340 may include: a predetermined driving device 342 that may be fixed to the second side frame 230; and a predetermined detaching plate 350 which can be displaced by the driving means 342. The driving means 342 and the detaching plate 350 may be connected by a predetermined shaft 344.

As shown in fig. 8, the take-down plate 350 is positioned between the holder 310 and the dispenser module 500. Referring to fig. 12, the disassembly plate 350 has a plate body 352, and a removal line formed by three disassembly holes 354a, 354b, and 355 aligned in a row is formed on the plate body 352. The removal lines are formed in a number corresponding to the number of mounting channels 312 formed in the holder 310. Two detachment holes 354a and 354b of the removal line are formed in a manner connected to each other and located between the holder 310 and the dispenser module 500 to pass a punch arm 552 and a suction tube arm 554, which will be described later, respectively. The acquisition arm 556 is formed by removing a detachment hole 355 formed separately on the wire.

Each of the disassembly holes 354a, 354b, and 355 may have a recess 356 recessed to one side. Accordingly, in a state where the dispensing tip 20 fastened to the collection arm 556 and the washing tip 30 fastened to the pipette arm 554 are located in the respective detachment holes 354a, 354b, 355, the detachment plate 350 is displaced in the left-side horizontal direction so that the collection arm 556 is located in the recess 356, at which time a part of the upper end of the dispensing tip 20 is located below the recess of the plate, and when the collection arm or the pipette arm is moved upward, the dispensing tip 20 fastened to the collection arm 556 or a part of the upper end of the washing tip 30 fastened to the pipette arm 554 is subjected to a force, so as to be detachable from the respective arms.

The detachment hole 355 is larger in area than the upper end of the dispensing tip 20 or the wash tip 30 so that the collection arm mounted with the dispensing tip or the pipette arm mounted with the wash tip can pass through the detachment hole. Preferably, the radius of the recess 356 is greater than the radius of the collection or pipette arm so that the collection or pipette arm can be mounted to the recess. Preferably, the area of the recess 356 is smaller than the area of the upper end of the dispensing tip or the washing tip so that the upper end of the dispensing tip or the washing tip can be caught in the protruding portion, however, the shape of the recess 356 is not critical as long as the dispensing tip or the washing tip can be separated from the collection arm or the pipette arm.

The reaction occurring in the cuvette 10 used in the device according to one embodiment of the invention requires at least two incubation processes from the start to the detection. Since the detachment module 340 is provided on the apparatus according to an embodiment of the present invention, there is an advantage in that only one dispensing tip and one washing tip can be used in one cuvette and reactions in other cuvettes mounted on other mounting channels 312 can be prepared during the incubation time.

Specifically, in order to dispense/mix reagents into cuvettes provided in the second mounting channel during a first incubation time during which an immunoreaction occurs in cuvettes mounted in the first mounting channel 312, the dispensing tip 20 and the washing tip 30 used in the first channel are temporarily kept in the respective positions 21, 32 of the first cuvettes, and after the first incubation time has elapsed, the dispensing tip 20 and the washing tip 30 temporarily kept can be reused. That is, when the module 340 is not disassembled, the dispensing tip 20 or the washing tip 30 used in the first mounting channel cannot be reused, and it is necessary to re-mount and perform the subsequent processes after the first incubation after discarding, and therefore, at least two dispensing tips 20 and two washing tips 30 are required for each cuvette disposed in the mounting channel. However, in the present invention, since the detachment module 340 is provided, there is an advantage in that the inspection process can be performed with only one dispensing tip 20 and one washing tip 30 per cuvette.

An apparatus according to one embodiment of the invention may include a standard block 360. The standard block 360 is fixed to the holder 310 so as to be integrally displaced with the holder 310, and may be located at the rear of the holder 310. Preferably, the standard block 360 may be positioned behind at least one of the inspection holes 314.

The standard block 360 has a predetermined optical hole 362 penetrating in the up-down direction, and a predetermined optical device capable of optical detection or capture may be provided on the optical hole 362.

In one embodiment of the present invention, standard block 360 includes optics. In one embodiment of the present invention, the standard block 360 includes an optical device loaded with a fluorescence measurement standard having a predetermined fluorescence value. The fluorescence measurement standard substance may use a substance having appropriate excitation and emission wavelengths depending on the type of fluorescence detected in the reaction product. In one embodiment of the present invention, 4-Methylumbelliferone sodium salt (4-Methylumbelliferone sodium salt) having an excitation (excitation) wavelength of 360nm and an emission (emission) wavelength of 450nm is used, but is not limited thereto.

In another embodiment of the present invention, the optics included in the standard block 360 are loaded with visible color (visible color) absorbance measurement standards. The absorbance measurement standard substance may be appropriately selected according to the absorbance region of the visible color detected in the reaction product, and in one embodiment of the present invention, glass (glass plate), plastic (plastic plate), gel (gel), an appropriate liquid phase solution, or the like is used, but is not limited thereto.

In the optical analysis, when measuring the fluorescence or absorbance value of the reaction product after the completion of the reaction, the standard fluorescence or absorbance loaded on the standard block 360 is scanned and then the signal value of the reaction product is measured and expressed by a ratio. This is to eliminate instrument to instrument variation, calculate the ratio to the measured value using the standard substance, and compare the ratio to the data of the built-in master calibration chart to accurately calculate the concentration of the analyte in the sample.

When measuring fluorescence or absorbance signals, the absolute value of the fluorescence value is usually different from device to device. Therefore, when the concentration is calculated using the absolute value of fluorescence, there may be a problem that an error is generated due to the equipment. Therefore, as one embodiment of the present invention, when the ratio of the standard substance to the measured value of the standard block is used, the error of the measured value between the devices is reduced, and the accuracy and reproducibility are improved.

An apparatus according to still another embodiment of the present invention may not include the standard block 360, or may not use the standard block 360 even if the standard block 360 is included. For example, when the signal detected in the reaction product is chemiluminescence, the standard block may not be included, or even if the standard block is included, the standard block may not be used. In this case, the device includes a light detector such as a photomultiplier tube (PMT), an avalanche photodiode (avalanche photodiode), and may further include a shutter implemented in hardware or software as a means for measuring the light amount during a predetermined time to measure the relative light amount, whereby a deviation of a detection signal between the means for comparing may be used to correct it.

The holder 310 may be displaced in the front-rear direction when the holder driving part 320 is operated. At this time, when the holder 310 moves backward by a predetermined distance, the standard block 360 fixed to the holder 310 is positioned on an optical discriminator 410 described later. Thus, the optical interpreter 410 can capture the fluorescence signal of the standard block 360.

Further, when the holder 310 moves rearward to the rearmost end, the rear lower portion of the holder 310 is positioned on an optical interpretation module 400 described later. Therefore, when the holder 310 is moved rearward to the rearmost end in a state where the cuvette 10 is mounted on the mounting passage 312 of the holder 310, the lower portion of the detection chamber 16 disposed rearward of the cuvette 10 may be exposed to the optical interpretation module 400 through the inspection hole 314.

Since the displacement of the holder 310 is guided by the holder guide 330, the displacement can be stably performed without shaking. In particular, since the driving part 320 of the transmission belt type fastener is provided, vibration and foreign matter caused by friction generated during movement can be prevented, and more accurate inspection can be performed compared to the gear type.

The optical interpretation module 400 is explained in detail below.

The optical interpretation module 400 is used to measure the signal of the reaction product within the cuvette 10. Preferably, the optical interpretation module 400 may include an optical interpreter 410, an interpreter driving part 420, and an interpreter guiding part 430.

The optical analysis is performed by the optical interpretation module 400. Such optical analysis includes measuring the fluorescence signal, visible colour and chemiluminescence of the reaction product, and reference may be made to the above for the definition of each signal.

Optical interpreter 410 may be positioned below holder 310 as holder 310 is moved toward the back end. Therefore, when the holder 310 is moved backward in a state where the cuvette 10 is accommodated in the holder 310, the detection chamber 16 of the cuvette 10 is located on the optical discriminator 410. Thus, the fluorescence values of the reaction products within the detection chamber 16 may be measured by the optical reader 410.

The optical interpreter 410 interprets the signal of the reaction product of the detection chamber 16 of the cuvette 10 so that a specific target analyte included in the sample can be analyzed qualitatively and/or quantitatively.

In one embodiment of the invention, the optical interpreter 410 of the optical interpretation module detects the fluorescent signal. An embodiment according to the present invention may be configured such that light of a specific wavelength is irradiated and the emitted light is interpreted according to the type of fluorescent substance used for detecting an analyte.

For example, the optical interpreter 410 may include a structure as shown in fig. 16 a. To analyze reaction product 650, optical interpreter 410 may include a light source 610, a collimating lens 620, a beam splitter 630, a focusing lens 640a, a filter 660a, a focusing lens 670a, and a light detector 680 a.

Within the optical discriminator 410, a light source 610 capable of sufficiently exciting a fluorescent substance, i.e., a predetermined light emitting element, may be provided to measure the fluorescent signal whose output can be adjusted. Examples of such Light Emitting elements include Xenon (Xenon) lamps, Ultraviolet (UV) lasers or Light Emitting Diodes (LEDs). In one embodiment of the invention, LEDs are used. Compared with Xenon lamps, UV lasers, etc., LEDs are inexpensive and equipment can be miniaturized. In one embodiment of the present invention, when an LED is used, a feedback circuit is built in to stabilize the temperature and the power supply portion, and the diffusion type LED is caused to emit light in parallel by using two pin holes.

In particular, as described above, before the fluorescence value is measured, light is irradiated to the standard block 360, and the increment (gain) is automatically adjusted by the amount of captured fluorescence, so that the output of the light emitting element can be adjusted to a predetermined value to calculate an accurate concentration.

On the other hand, the optical discriminator 410 may have more than two light sources, and each light source may generate light having a different wavelength. In addition, fluorescence at different wavelengths can be measured separately. Therefore, the application range of the diagnostic test method can be expanded, and the sensitivity is more excellent.

In addition, the optical discriminator 410 may have a barcode scanner function, and thus, when a predetermined barcode is provided on the cuvette 10, predetermined signals, information, and the like may be exchanged by the corresponding barcode.

In yet another embodiment of the invention, the optical interpreter 410 of the optical interpretation module measures the absorbance of the visible color of the reaction product 650. The absorbance may be measured by irradiating light to the reaction product according to the kind of the substance used in the detection of the analyte according to one embodiment of the present invention. On the other hand, the optical discriminator 410 includes a light source therein, which can be adjusted in output and can emit an absorption band suitable for the absorbance measurement of the visible color. Examples of such light emitting elements may include, but are not limited to, lamps including absorption bands such as white light sources, LEDs, lasers, and the like.

In yet another embodiment of the invention, the optical interpreter 410 measures the chemiluminescent signal of the reaction product. The optical discriminator 410 may be configured such that the kind of chemiluminescent substance used in the detection of the analyte according to one embodiment of the present invention detects the emitted light, and is composed of a lens for capturing light and a photodetector since the luminous intensity of light is measured over a period of time.

For example, the optical interpreter 410 may include a structure as shown in fig. 16 b. To analyze reaction product 650, optical interpreter 410 includes focusing lenses 640b, 670b and photodetector 680 b. For more accurate analysis, the optical interpreter 410 may further include a filter 660b, in which case the optical interpreter 410 does not include a light emitting element or light source, but rather a light detector 680b, such as a photomultiplier tube or avalanche photodiode.

In addition, in order to measure the relative light amount, a shutter implemented in hardware or software may be provided as a means for measuring the light amount during a predetermined time, and thus it is possible to correct it by comparing the deviation of the detection signal between the means.

The discriminator driving part 420 is provided inside the casing 100, and by moving the optical discriminator 410, the optical discriminator 410 is located on any one of the cuvettes 10 so that the sample of the cuvette 10 can be inspected. That is, the discriminator driving section 420 may move the position of the optical discriminator 410 in accordance with the inspection hole 314 of the holder 310.

For example, the discriminator driving section 420 may include a predetermined driving motor 422 that can move the optical discriminator 410 left and right, a driven wheel 424, and a predetermined support that connects the driven wheel 424 and the optical discriminator 410. Accordingly, the optical discriminator 410 can be moved according to the operation of the driving motor.

The discriminator guide 430 is provided to guide the left-right directional displacement of the optical discriminator 410. The interpreter guide 430 may include a predetermined guide rail and a predetermined guide portion that is guided along the guide rail and fixed to the optical interpreter. Therefore, the right-left direction movement of the optical discriminator can be accurately guided in one direction.

As described above, at this time, when the holder 310 moves backward by a predetermined distance, the standard block 360 at the lower rear portion of the holder 310 is positioned on the optical interpretation instrument 410 of the optical interpretation module 400. Therefore, first, the optical interpretation module 400 detects the fluorescence signal captured in the standard block 360 as standard fluorescence.

Subsequently, when the holder 310 is moved backward to the rearmost end in a state where the cuvette 10 is mounted on the mounting passage 312 of the holder 310, the lower portion of the detection chamber 16 disposed at the rear of the cuvette 10 may be exposed to the optical discriminator 410 through the inspection hole 314 to perform optical measurement.

At this time, as described above, the ratio of the fluorescence signal captured by the proof mass 360 to the fluorescence signal captured by the detection chamber 16 is expressed. The optical interpretation module 400 may have a predetermined algorithm and a predetermined repeated measurement algorithm such that the above ratio is compared to the data of the built-in master calibration chart to calculate the analyte concentration in the sample.

As described above, the measurement is performed in the form of comparing the fluorescence value of the standard fluorescence loaded on the standard block 360 with the fluorescence value of the sample, so that accurate measurement can be performed. That is, according to the general prior art, fluorescence values vary from device to device, and in order to reduce such variations, a calibration process that reduces variations between instruments is mostly required at the QC step. However, despite this process, it is difficult to completely eliminate this difference due to changes in instruments or reagents. However, in the present invention, since the standard fluorescence loaded on the standard block 360 serves as a reference, the above-mentioned problems can be solved.

Hereinafter, the dispenser module 500 is explained. Fig. 14a and 14b are exploded views each showing the structure of the dispenser module 500 in the automated liquid-phase immunoassay device 1 according to an embodiment of the present invention from different directions.

The dispenser module 500 is a module provided for dispensing, dispensing and washing a sample, a reagent and/or a reaction substance.

The dispenser module 500 includes a drive unit 502, a dispenser unit 504, and a pump unit 506.

First, the driving unit 502 will be explained.

The driving unit 502 is used to move the dispenser unit 504 left and right in the horizontal direction. Accordingly, the dispenser unit 504 is horizontally moved by the driving unit 502 so that the dispenser unit 504 can be located in a specific chamber on any one of the cuvettes 10 disposed side by side below the driving unit.

The driving unit 502 may include a fixed body 510 and left and right horizontal driving parts 520.

The fixing body 510 may have a predetermined area and may extend long in the left-right direction. The fixing body 510 may include a front body 512 extending in a left-right direction and a side body 514 disposed at one side of the front body 512 to fix the pump unit 506.

The left-right driving part 520 is provided on the fixing body 510, and the left-right driving part 520 is a driving device that moves the later-described dispenser unit 504 left and right in the horizontal direction. The left and right driving part 520 may include a predetermined driving motor generating power and a predetermined moving bracket displaceable left and right by the driving motor. In addition, the left and right driving part 520 may be provided with a predetermined guide device 530 that can guide the displacement of the moving bracket. In addition, the left and right driving part 520 may include a predetermined driven wheel member to transmit power.

Next, the dispenser unit 504 will be explained. The dispenser unit 504 may include a left-right moving body 540, an up-down moving body 542, an up-down driving part 544, and an arm unit 550.

The left and right moving bodies 540 are connected to the left and right driving parts 520. As described above, the left-right driving part 520 includes a predetermined moving bracket, and the left-right moving body 540 is connected to the moving bracket so as to be displaceable left and right in the horizontal direction.

The up-down moving body 542 is provided in front of the left-right moving body 540. The vertical moving body can be vertically displaced by the vertical driving unit 544.

The up-down driving unit 544 is provided on the left-right moving body 540, and the up-down driving unit 544 is a driving device that moves the up-down moving body 542 in the up-down direction. The up-down driving part 544 may further include a predetermined driving motor generating power and a predetermined moving bracket that can be displaced left and right by the driving motor. The up-down driving unit 544 may be provided with a predetermined guide device 546 capable of guiding the displacement of the movable bracket in the up-down direction. In addition, the up-down driving part 544 may include a predetermined driven wheel member that transmits power.

The arm unit 550 is a member that can move up and down by the up-and-down driving unit 544 and can move left and right by the driving unit 502. The arm unit 550 may include a punch arm 552, a collection arm 556, and a suction pipe arm 554 connected to the upper and lower moving bodies 542 and extending downward at positions spaced apart from each other in the horizontal direction. Thus, the arm unit 550 may constitute an integrated module in which the perforating arm 552, the collecting arm 556 and the pipette arm 554 are integrally formed.

The punch arm 552 is provided at a lower end thereof with a punch tip 553, the punch arm 552 is a member that penetrates the sealing lid of the cuvette 10 to be opened, and the punch arm 552 penetrates a sealing portion covering the corresponding chamber of the cuvette 10.

The straw arm 554 is vertically penetrated and has a hollow upper and lower portions 555. The pipette arm 554 has an outer diameter that can be plunged and inserted into the insertion hole of the wash tip 30.

The collection arm 556 is provided so that the dispensing tip 20 can be fixed at the lower end. The collection arm 556 may have an outer diameter that can be plunged and inserted into the dispensing tip 20.

Preferably, the punch arm 552, the pipette arm 554, and the collection arm 556 may be arranged in a row in the front-rear direction.

The washing unit 560 includes a driving motor 562 and a magnetic beam 564.

The driving motor 562 is fixed to the upper and lower moving bodies 542, and is connected to the magnetic flux 564 so that the magnetic flux 564 can be displaced in the upper and lower directions. On the other hand, it is not necessary to limit the present invention to the driving motor 562, and a predetermined driving device capable of vertically displacing the magnetic flux 564 may be provided.

The magnetic beam 564 is provided in a strip shape extending in the up-down direction and is provided in the upper and lower hollows 555 of the pipette arm 554. The magnetic beam 564 has magnetism and can be displaced in the up-down direction by the driving motor 562, so that a magnetic bead separation technique (Mag-eXtraction) using magnetic separation of unreacted substances can be performed.

The pump unit 506 is fixed to the side body 514 of the drive unit 502. The pump unit 506 is connected to the collection arm 556 of the dispenser unit 504 through a predetermined tube (not shown), so that the pump unit 506 functions to provide a suction force or a discharge force when the dispensing tip 20 is inserted into the chamber of the cuvette 10 in a state of being connected to the collection arm 556. Specifically, when the cuvette 10 is positioned at a specific position by the cuvette module 300 and the dispensing tip 20 is put into the chamber in a state where the dispensing tip 20 is positioned on the chamber of the cuvette 10 by the driving unit 502, the pump unit 506 may supply a suction force or a discharge force to the dispensing tip 20. Preferably, the pump unit 506 has a motor 570 that can control the rotational micro-steps, and can be configured to accurately control the amount of the sample when the sample, the reagent, or the reaction product is aspirated or discharged from the dispensing tip 20.

Fig. 15a is a configuration diagram showing a schematic structure of a dispenser module in an automated liquid-phase immunoassay apparatus according to another embodiment of the present invention.

The dispenser module includes a moving body 541, a moving body driving section 543, and a control section 600. The control unit 600 may control the moving body driving unit 543 to move the moving body 541 to a desired position.

A punching arm 552, a pipette arm 554, and a collection arm 556 are fixed to the moving body 541. Therefore, the punching arm, the straw arm, and the collection arm are integrally moved by the movement of the moving body.

The lower portion of the punch arm 552 is provided with a punch tip 553. When the punch arm penetrates the seal of the cuvette at the lower portion of the punch arm, the pipette arm and the collection arm, which are fixed to the moving body 541 together with the punch arm and move integrally, should not interfere with the cuvette at the lower portion. That is, the length B from the lower portion of the moving body to the lower portion of the punching arm 552 should be longer than the length a of the pipette arm and the collection arm. The appropriate length can be set so that the pipette arm and the collection arm do not contact the cuvette even though the punch arm is maximally lowered to penetrate the cuvette seal.

When the pipette arm 554 with the wash tip 30 attached thereto or the collection arm 556 with the dispense tip 20 attached thereto is operated with a cuvette, the punch arm 552 should not interfere with the lower cuvette. Therefore, the length B from the lower portion of the moving body to the lower portion of the punch arm 552 should be shorter than the length C from the lower portion of the moving body to the end portion of the wash tip mounted on the pipette arm or the end portion of the dispensing tip mounted on the collection arm. That is, the height of the wash tip and the dispense tip should be greater than the sum of the length of the punch arm and the depth of each chamber within the cuvette. Each tip may be set to an appropriate length in consideration of a mounting position of each tip to each arm and a smooth operation distance in each chamber.

The collection arm 556 may fixedly mount the dispensing tip 20 at a lower portion. A hollow 557 penetrating vertically is provided in the collection arm. The hollow of the acquisition arm is connected to a pump unit 506 through a tube 507. The pump unit 506 can provide a suction force and a discharge force to the dispensing tip through the hollows of the tube and the collection arm.

The pipette arm 554 may fixedly mount the wash tip 30 at a lower portion. The interior of the straw arm is provided with an upper hollow 555 which is communicated up and down. A magnetic beam 564, which can move up and down, is located in the hollow of the pipette arm. The drive motor 562 is provided to move the magnetic flux up and down. Preferably, the driving motor 562 is fixed to the moving body to enable the relative movement of the magnetic flux with respect to the suction pipe arm fixed to the moving body.

The drive motor and the magnetic flux may be connected by a linear actuator using a ball screw or the like, a reduction gear using a gear coupling, a rack and pinion gear, or the like.

FIG. 15b is an enlarged view illustrating in detail a washing tip portion of a dispenser module in an automated liquid-phase immunoassay device according to another embodiment of the present invention

Magnetic beam 564 is disposed within upper and lower hollows 555 of pipette arm 554. The magnetic beam 564 may be provided with a permanent magnet 565 at a lower portion thereof, which is an end opposite to a portion connected to the driving motor 562. Preferably, the permanent magnet 565 has the same cross-sectional area as the shape of the attached magnetic beam. When the magnetic flux is cylindrical in shape, a cylindrical permanent magnet having the same diameter may be used. When the magnetic beam 564 is lowered by the driving motor 562, the permanent magnet may be disposed inside the wash tip 30 inserted into the pipette arm 554.

Preferably, the diameter of the permanent magnet 565 is 2mm to 8mm when considering the chamber size of the cuvette. When the length of the permanent magnet is 5mm or more, the magnetic beads may be captured, however, it is preferable to use a permanent magnet of 10mm or more in order to collect the magnetic beads required for measurement within 1 minute. More preferably, when a permanent magnet of 30mm or more is used, sufficient magnetic beads can be collected within 40 seconds. The shape of the permanent magnet may be selected and used according to purposes, and various shapes such as a circle, a quadrangle, and an ellipse.

Hereinafter, the operation of the automated liquid-phase immunoassay device 1 according to an embodiment of the present invention will be described with reference to fig. 17 to 20.

Fig. 17 is a flowchart illustrating an overall flow of an automated liquid-phase immunoassay method according to an embodiment of the present invention.

First, the cuvette 10 is accommodated in the mounting passage 312 of the holder 310 of the apparatus 1 (S710). At this time, the dispensing tip 20 and the washing tip 30 are mounted in the dispensing tip insertion hole 21 and the washing tip insertion hole 31 formed in the cuvette (S720). The dispensing tip 20 and wash tip 30 may be mounted before or after the cuvette 10 is received in the mounting channel 312. Then, the holder 310 is moved backward by a start command of the device (S730).

Subsequently, the dispenser module 500 is operated to punch and open a sealing film (not shown) of the cuvette 10 (S740). During the punching process, a punching arm 552 is used. The punching process is explained as follows, first, the punch arm 552 is positioned on the cuvette 10 by the driving unit, and then, the punch arm 552 is moved up and down by the up-down driving unit 544 to punch the sealing film of the cuvette 10. In this process, the cuvette module 300 is operated to move the cuvette 10 forward or backward so that a plurality of chambers provided in the cuvette 10 can be perforated.

Subsequently, when the punching is completed, the cuvette module 300 and the dispenser module 500 are operated such that the collection arm 556 is positioned on the dispensing tip 20 fixed to the cuvette 10. Subsequently, the collection arm 556 is lowered, and the dispensing tip 20 is inserted and fixed to the lower portion of the collection arm 556 (S750). Thereafter, the sample and/or the reagent is dispensed and dispensed using the dispensing tip 20 (S760).

At this time, the detailed description of the dispensing process is as follows. First, the movable body 541 to which the collection arm is fixed is moved to drop the dispensing tip into the sample solution. Then, the hollow pump unit 506 connected to the collection arm is operated to apply a suction force to the dispensing tip 20 to collect a sample from the sample chamber. Next, the moving body driving part 543 is driven to move the collection arm fixed to the moving body to the reaction chamber. At this time, the sample attached to the dispensing tip of the collection arm is also moved to the reaction chamber. That is, the collected sample may be moved to the reaction chamber. Then, the pump unit 506 is operated to apply a discharge force to the dispensing tip 20, and the sample is discharged to the reaction chamber to complete dispensing.

In this process, as described in the above-described punching process, the cuvette 10 can be moved forward or backward by the cuvette module 300, and the dispensing tip 20 can be moved upward and downward by the vertical driving unit 544. At the same time, the pump unit 506 is operated to effect dispensing and dispensing through the dispensing tip 20. In addition, by operating the pump unit 506, mixing of the sample and/or reagent can be achieved during dispensing and dispensing, and a desired reaction occurs in the reaction chamber 14 of the cuvette.

Thus, the reaction process occurring in the cuvette 10 includes a plurality of steps, and each cuvette requires at least two incubation times (S770). The incubation may be performed by applying power to the hot plate 316 of the holder 310 holding the sample to maintain the sample dispensed in the reaction chamber at a constant temperature.

Therefore, in order to start the reaction of the second cuvette during the first incubation time, the used dispensing tip 20 in the first cuvette is detached and positioned in the dispensing tip insertion hole 21 of the first cuvette by the detaching plate 350. After the first incubation time is complete, the used dispensing tip 20 in the first cuvette is reused for the next reaction in the first cuvette.

The incubated sample is subjected to a washing process (S780). After the washing is completed, the sample containing the magnetic beads from which the impurities are removed is moved to a detection chamber, subjected to an optical inspection process, and used for analysis (S790).

Fig. 18 is a flowchart illustrating a washing process of an automated liquid-phase immunoassay method according to an embodiment of the present invention.

The washing process includes a step of trapping magnetic beads, a step of moving and putting the trapped magnetic beads into a washing solution, and a step of removing impurities.

That is, first, the washing tip into which the magnetic beam is inserted is put into the sample solution containing the magnetic beads to trap the magnetic beads in the sample solution on the surface of the washing tip (S820 to S845). Next, the washing tip with the magnetic beads trapped on the surface thereof is moved to the washing solution in a state where the magnetic beam is inserted, and the washing solution is poured into the washing solution (S850). Then, a driving motor connected to the magnetic beam is driven to move the magnetic beam upward and move the washing tip up and down several times to disperse the magnetic beads trapped at the washing tip into the washing solution (S860). Thereafter, a magnetic beam is inserted into the washing tip to capture the magnetic beads in the washing solution again (S870). When less than the predetermined number of washing times, the washing tip having the magnetic beads trapped therein may be moved to a new washing chamber to repeat the above-described process until the predetermined number of washing times is reached (S880). After the washing is completed, the optical measurement may be performed by moving to the detection chamber (S890).

The washing process may be performed by various methods, and the position of the washing tip may move the magnetic beam up and down several times by driving a driving motor connected to the magnetic beam in a fixed state, thereby repeatedly dispersing and trapping the magnetic beads at the washing tip in the washing solution to remove impurities not bound to the magnetic beads.

Meanwhile, the step of trapping the magnetic beads in the sample solution can be classified as follows. First, the washing tip 30 is fixed to the lower portion of the suction pipe arm 554 having a hollow therein penetrating up and down (S820). Then, the movable body 541 to which the pipette arm is fixed is lowered, and the sample solution containing magnetic beads is poured into the washing tip (S830). Next, the driving motor 562 fixed to the moving body is driven to insert the hollow magnetic flux 564 located at the pipette arm into the washing tip at the lower portion (S840). Thereafter, the moving body driving part 543 may integrally move the moving body to which the pipette arm is fixed and the magnetic flux so that the washing tip into which the magnetic flux is inserted moves within the sample solution containing the magnetic beads (S845).

The above process is described in detail as follows. When the dispensing, and reaction of the sample and the reagent are completed, the dispensing tip 20 is detached from the collection arm 556 by the detachment plate 350 (S810). Subsequently, the wash tip 30 is inserted to the pipette arm 554 (S820). The washing tip 30 is thrown into the reaction chamber 14 (S830), and then the magnetic beam 564 is thrown into the washing tip 30, thereby trapping the magnetic beads in the reaction chamber 14 on the surface of the washing tip 30 (S840). At this time, the reactant bound to the magnetic beads is captured together. To more effectively trap the magnetic beads, the washing tip and the magnetic beam may be moved together within the sample solution (S845). After moving the washing tip 30 and moving it into the washing chamber 15 in this state (S850), and raising the magnetic beam by the driving motor 562 to separate the magnetic beam 564 from the washing tip 30, the magnetic beads trapped at the washing tip 30 are dispersed in the washing chamber 15 (S860). At this time, the washing tip may be moved up and down several times so that the magnetic beads trapped at the washing tip are well dispersed in the washing solution. When the magnetic beam 564 descends again and moves to the washing tip 30 side, the magnetic beads are trapped to the washing tip 30 again (S870). The rising and falling of the magnetic beam performs washing a predetermined number of times (S880). Non-magnetic impurities can be removed in the washing chamber by the reciprocating motion of the magnetic beads as the magnetic beam rises and falls. After the sample washing is finished, the reaction product is moved to the detection chamber 16 (S890).

Fig. 22 is a photograph showing the effect of a permanent magnet on a sample containing magnetic beads used in an automated liquid-phase immunoassay method according to an embodiment of the present invention. For visualization, a permanent magnet was placed under a solution containing a sample of high-concentration magnetic beads, and the influence of the magnetic beads was observed with the passage of time, and the magnetic beads were dispersed in a 0-second photograph, and thus the whole was displayed as a yellow solution. Over time, the beads were pulled to the permanent magnet side of the bottom, so the solution gradually became transparent and the yellow color of the bottom appeared darker. After about 50 seconds, it was confirmed that the magnetic beads were sufficiently pulled to the permanent magnet at the bottom.

Fig. 23 is a graph showing the magnetic field strength using a permanent magnet in an automated liquid phase immunoassay method according to an embodiment of the present invention. The permanent magnet used for the simulation was a cylindrical magnet having a diameter of 8mm and a thickness of 4mm, and was a result of the simulation for the case of 3700 Gauss (Gauss) magnetic field and 846kA/m coercive force.

Fig. 23 (a) shows the magnetic field intensity according to the distance from the magnet, and it is known that the magnetic field intensity decreases as the distance from the magnet increases. That is, the magnetic field strength of 200mT is exhibited at 2mm from the magnet, and when it exceeds 2mm, the magnetic field strength becomes weak, and thus the force pulling the magnetic beads is also reduced. The maximum height of the solution was about 10mm, and the magnetic force at a position 10mm from the magnet was almost close to 0.

Fig. 23 (b) shows the magnetic field intensity according to the position around the magnet, and it can be seen that when the ranges of the turquoise and sky-blue portions outside the magnet are observed in addition to the inside of the magnet, it is known that a stronger magnetic field is widely distributed in the up-down direction of the magnet than in the circumferential direction of the magnet. As a result, when the cylindrical permanent magnet is used, the magnetic beads can be influenced more by the upper and lower magnetic fields of the magnet than by the cylindrical side surface portion of the cylindrical permanent magnet, and the magnetic beads can be trapped or washed more quickly.

Fig. 24 is a photograph showing the position of a washing tip used in an automated liquid-phase immunoassay method according to an embodiment of the present invention. In these photographs, high concentrations of magnetic beads were added for visualization.

As shown in fig. 24 (a), in the automated immunoassay method according to one embodiment of the present invention, the washing tip may be removed after a predetermined time after it is kept in contact with the bottom of the washing chamber. In this case, as shown in (b), most of the magnetic beads can be captured after 45 seconds.

As shown in (c) of fig. 24, in the automated immunoassay method according to another embodiment of the present invention, the washing tip may be removed after a predetermined time after it is kept in the middle of the washing chamber solution. In this case, the magnetic beads can be captured almost completely after 45 seconds as shown in (d). This is because, when the thickness of the magnet used is small, the region where the strong magnetic field spreads is larger in the upper and lower portions of the magnet, and therefore, more magnetic beads can be pulled in a faster time.

An automated immunoassay method according to yet another embodiment of the present invention is to change the position of the washing tip while capturing and washing the magnetic beads. That is, since the magnetic force of the permanent magnet varies with distance, when the magnetic beads are collected, the magnetic beads can be collected more efficiently when the washing tip including the permanent magnet is moved up and down with reference to the middle of the solution including the magnetic beads. The up-down position can select the range of the magnetic beads favorable for trapping from the distance between the surface of the solution and the bottom of the solution and move.

In addition, considering the time when the magnetic beads are pulled onto the permanent magnet for the trapping, a stepwise up-and-down movement may be used, which stops the washing tip at a predetermined position in the solution containing the magnetic beads for a predetermined time until the magnetic beads are trapped, and then slightly advances the position of the washing tip in the solution again and stops again for a predetermined time after the predetermined time elapses. With such stepwise lifting movement, i.e. with a discriminating operation, the magnetic beads can be more completely trapped in a short time. The distinguishing operation may determine the distance of the moving section and the time of the stop in consideration of the washing time.

For example, when the washing time is set to 40 seconds or less and the moving interval of the washing tip is set to 4mm, the washing tip with the magnetic beam mounted thereon is stopped at the position dropped into the initial solution for 5 seconds, then moved downward for 1mm and then stopped for 5 seconds, and then lowered again for 1mm and then stopped for 5 seconds, and this process is repeated to move the washing tip in the solution for 4 mm. After moving to the lower part of the solution, the washing tip may be raised again by 1mm, stopped for 5 seconds, and then raised again by 1 mm. This allows for complete capture of the magnetic beads in a shorter time, and thus allows for a faster, more complete washing process.

According to the embodiment of the present invention, in order to capture magnetic beads, the washing tip into which the magnetic beam having the permanent magnet is inserted may be stopped at an intermediate position in the sample solution containing the magnetic beads to capture the magnetic beads. Preferably, the washing tip into which the magnetic beam is inserted and the magnetic beam are integrally moved up and down together to trap the magnetic beads in the sample solution containing the magnetic beads. More preferably, when moving the washing tip and the magnetic beam up and down together in the sample solution containing the magnetic beads, a stepwise up-and-down movement that stops for a predetermined time after moving for a predetermined distance may be performed to more rapidly and completely capture the magnetic beads.

This method of trapping magnetic beads on the washing tip can be applied not only to the step of trapping magnetic beads in the sample solution (S845) but also to the step of trapping magnetic beads in the washing solution (S870) in the same manner.

Meanwhile, in the above process, the separation of the dispensing tip 20 and the washing tip 30 may be achieved by the detachment module 340. That is, after the dispensing tip 20 or the washing tip 30 is positioned in the detaching hole 354 of the detaching plate 350, the detaching plate 350 is moved to move the dispensing tip and the washing tip to the recess 356, and when the collection arm or the pipette arm is moved upward, the upper end of a portion of the tip mounted on the collection arm and the pipette arm is caught in the recess 356, so that the dispensing tip 20 or the washing tip 30 can be separated from the collection arm 556 or the pipette arm 554.

The separation of the dispensing tip 20 and the wash tip 30 can be achieved in the same way. The separation of the washing tip 30 will be specifically described, and can be performed according to the following sequence. The knock-out plate 350 is positioned between the holder 310 and the dispenser module 500. First, the detaching hole 354 of the detaching plate 350 is located on the washing tip inserting hole 21. That is, a detaching plate 350 having a detaching hole 355 formed with a recess 356 is provided below the pipette arm 554 to which the washing tip 30 is mounted. Subsequently, the pipette arm mounted with the wash tip passes through the detachment hole 355 having a larger area than the upper end of the wash tip, so that the wash tip 30 is positioned in the wash tip insertion hole 21. Thereafter, the detaching plate 350 is lapped on the washing tip 30. That is, the recess 356 of the detaching plate is disposed above the upper end of the washing tip. In this state, the suction pipe arm is moved upward relative to the detaching plate so that the upper portion of the washing tip 30 is caught in the recess of the detaching plate, whereby the washing tip can be separated from the suction pipe arm. That is, the dispenser module 500 is raised to catch the wash tip 30 on the detaching plate 350, so that the wash tip 30 is separated from the pipette arm 554 and placed in the wash tip insertion hole 21.

Therefore, since the separated dispensing tip 20 and the wash tip 30 remain in the wash tip binding well 21 and the dispensing tip binding well 31, respectively, after separating the dispensing tip 20 and the wash tip 30, they are not mixed with other samples, so that there is no need to wash the tips, and they can be repeatedly used for the reaction of the next step in the same cuvette.

Subsequently, when the reaction product moves into the detection chamber 16, the optical interpretation module 400 is operated to perform an optical inspection. At this time, the optical discriminator 410 is located below the detection chamber 16. In addition, as described above, since the detection chamber 16 has optical transparency, the optical reader 410 can perform optical inspection of the reactant inside.

Fig. 19 is a flowchart illustrating an optical inspection process using a standard block in an automated liquid-phase immunoassay method according to an embodiment of the present invention.

First, the standard block 360 at the rear lower portion of the holder 310 is disposed above the optical interpreter (S910). To this end, the holder 310 or the optical interpreter 410 may be moved so that the standard block 360 may be positioned above the optical interpreter 410. The optical interpreter 410 may first perform an optical inspection of the fluorescence measuring standard substance in the standard block 360 located behind the holder 310 to interpret the fluorescence signal of the standard substance (S920). Thereafter, the optical discriminator 410 is disposed below the detection chamber (S930). That is, the holder or optical reader is moved so that the detection chamber 16 can be positioned above the optical reader 410. The optical reader 410 performs an optical inspection of the sample in the detection chamber 16 through the inspection hole 314 penetrating the holder 310 having the detection chamber mounted thereon in the vertical direction and reads an optical signal emitted from the sample (S940). As described above, the signal captured from the standard block 360 may be used as a standard fluorescence value to correct a deviation between equipment (S950). That is, the signal obtained as a result of the optical inspection of the sample of the detection chamber and the signal obtained as a result of the optical inspection of the standard substance of the standard block are compared and the difference thereof is analyzed, so that the sample of the detection chamber can be corrected, and thus more accurate results can be obtained.

In addition, the apparatus according to an embodiment of the present invention may further include a chip insertion part (not shown) disposed on the housing and into which a chip containing analysis information is inserted. The chip inserted into the chip insertion portion is connected to the bar code of the cuvette. The barcode of the cuvette includes the substance to be analyzed (item) and batch (lot) information of the cuvette, and is linked to the chip. The chip includes a main calibration curve required in the concentration calculation of the analyte and information for driving the device according to the type of the analyte in the sample, so that the device can be driven according to the types of various analytes through linkage with the barcode for optimal examination. Therefore, various analytes can be easily examined by one device, and reproducibility and reliability of examination can also be improved. The bar code obtains information through a bar code scanner that scans the bar code.

The inspection process according to an embodiment of the present invention is explained in order as follows.

Here, a case where the cuvette 10 having the structure shown in fig. 6 is used will be described as an example. The cuvette 10 used in the examination carried out according to the present invention may have a structure as shown in fig. 6. Specifically, the cuvette 10 may be provided with a sample filling chamber 12, a buffer and dilution chamber 13, a reaction chamber 14, a washing chamber 15, and a detection chamber 16, the buffer and dilution chamber 13 including an MB buffer chamber 13a, a chamber 13b filled with a detection buffer (detection buffer) such as Alkaline phosphatase (ALP), a dilution buffer chamber 13c, and a dilution chamber 13d, and the washing chamber 15 may include a first washing chamber 15a and a second washing chamber 15 b.

Fig. 20 is a flowchart illustrating in detail a sample dispensing method of an automated liquid phase immunoassay method according to an embodiment of the present invention.

First, after the bar code is recognized, the sealing of the colorimetric dishes 10 are punched to be opened by the punching arms 552, respectively. Subsequently, the dispensing tip 20 is inserted and fixed on the collection arm 556. Subsequently, a predetermined volume of the washing liquid is collected from the first washing chamber 15a and dispensed to the MB buffer chamber 13a (S1010).

Subsequently, a predetermined diluent is collected from the dilution buffer chamber 13c and dispensed into the sample chamber 12(S1020), and a mixing process is performed (3 times). Subsequently, a predetermined volume of the diluted sample is collected and dispensed into the reaction chamber 14 (S1030). Subsequently, after the chamber 13b filled with the detection buffer is mixed, a predetermined volume of solution is collected and dispensed into the reaction chamber 14(S1040) and mixed (3 times). Subsequently, a first incubation process is performed during a predetermined time at a specific temperature (S1050). Subsequently, after the mixing of the MB buffer chamber 13a, a predetermined volume of the solution in the MB chamber 13a is collected and dispensed into the reaction chamber 14(S1060) and mixed, and then the dispensing tip 20 is detached using the detachment module 340 (S1070) and positioned at the dispensing tip insertion hole 21 of the cuvette in which the reaction is performed. In addition, a second incubation process is performed during a predetermined time at a specific temperature (S1080).

Subsequently, after the second incubation time elapses, a washing process is performed (S1090). The washing process is performed by first inserting the washing tip 30 into the pipette arm 554, and throwing the magnetic beam 564 into the pipette arm 554 to be thrown into the reaction chamber 14 during a predetermined time, and then into the first washing chamber 15a, and then moving the magnetic beam 564 up and down several times to perform washing. Subsequently, the magnetic beam 564 is thrown again into the pipette arm 554 and into the second washing chamber 15b, and then the magnetic beam 564 is moved up and down several times to perform washing. Subsequently, the magnetic beam 564 is again plunged into the pipette arm 554 and into the detection chamber 16, and then the wash tip 30 is removed.

Subsequently, after performing the third incubation process during a predetermined time, an optical measurement process is performed. The results (density, etc.) obtained by the optical measurement can be output to a display and a printer.

In addition, the reaction in another cuvette may be performed during the incubation. Fig. 21 is a timing diagram illustrating the operation of each cuvette when three cuvettes are included in the automated liquid phase immunoassay method according to one embodiment of the present invention. FIG. 21 shows the sequence of each step of dispensing, washing, incubating, etc. for three cuvettes.

Each step may be driven by being divided into step 1, step 2, step 3, step 4, step 5, step 6, etc., each step may be driven by dilution, collection, dispensing, mixing, washing, incubation, measurement, etc., and steps may be added or deleted according to purpose.

In order to drive and measure the three cuvettes simultaneously, each step should be driven separately from the other steps. Fig. 21 is an example of a method (protocol) for inspecting three cuvettes. The start and end points of each step are clearly distinguished from the operation of the other cuvette, and finally, the time required to perform measurements and experiments on all three cuvettes can be significantly reduced. In particular, when a preparation operation, a dispensing operation, or a washing operation is performed in another cuvette during the incubation period of one cuvette, a measurement time can be saved.

For example, when the inspection time of one cuvette is 20 minutes, it takes more than 60 minutes in total to inspect three cuvettes, however, when the above-described method is used, the inspection of three cuvettes can be completed within about 23 minutes using only one pump module, so that the time required for measurement and analysis can be reduced.

In order to perform a measurement process using such a plurality of cuvettes, a plurality of dispensing tips and washing tips are required to collect, dispense, and dilute the reagent of each cuvette to prevent contamination between cuvettes. The present invention is designed to include a knock-down plate formed with a recess so that the dispensing tip and the washing tip can be seated in the recesses of the respective cuvettes. After placing the tips for reagent collection, dispensing and washing in each cuvette back into the recesses of the corresponding cuvette, the dispensing tips and washing tips of the other cuvettes are mounted on respective holders so that a plurality of cuvettes can be inspected simultaneously in a short time without contamination of the cuvettes to each other

In the following embodiments, only a few examples of replacing the dispensing tip or the washing tip are described, however, it is preferable that, before the operation is performed on each cuvette, the dispensing tip or the washing tip used in the other cuvette is detached, and the dispensing tip or the washing tip of the cuvette to be operated is attached.

First, after preparation works such as sample dilution and addition of a reactant are performed in the first cuvette (S1111), the first incubation is started (S1112). After the same preparation work is performed on the waiting (S1120) second cuvette (S1121), the first incubation is started (S1122). Subsequently, when the first incubation of the first cuvette is ended, after performing the dispensing work of adding magnetic beads or the like (S1113), the second incubation is started (S1114). Thereafter, the tip 20 used for dispensing the sample in the first cuvette is detached from the collection arm 556 and mounted on the first cuvette.

Then, after the preparation work (S1131) is performed on the third cuvette waiting for the above-described period (S1130), the first incubation is started (S1132).

During the second incubation performed in the first cuvette, after the collection arm 556 mounts a dispensing tip to be used for dispensing the sample in the second cuvette, and the operation of adding a magnetic bead aliquot is also performed in the second cuvette (S1123), the second incubation is started (S1124). When the second incubation is initiated, the tip used in the dispensing of the sample in the second cuvette is detached from the collection arm 556.

Similarly, after the dispensing operation is also performed in the third cuvette (S1133), the operations may be sequentially performed so that the second incubation is started (S1134).

Then, during the incubation of the second and third cuvettes, the washing tip 30 to be used for washing the sample in the first cuvette is mounted at the pipette arm 554, and at the end of the second incubation of the first cuvette, a washing operation is performed (S1115), and the third incubation is started (S1116). During the third incubation period in the first cuvette, a washing operation is also performed in the second cuvette (S1125), and the third incubation is started (S1126). Similarly, a washing operation is also performed in the third cuvette (S1135), and a third incubation is started (S1136).

When the third incubation is completed in the first cuvette, measurement is performed (S1117). When the third incubation is also completed in the second cuvette, measurement is performed (S1127). Likewise, when the third incubation is also completed in the third cuvette, measurement may be performed (S1137).

Compared to the existing method, the automated liquid-phase immunofluorescence analysis device 1 according to one embodiment of the present invention can detect/interpret reaction products with high sensitivity and high specificity by way of sample dispensing and reaction, separation (purification) of reaction products by way of a washing module using Magnetic Beads (Magnetic Beads), and use of a liquid-phase sample optical system. In particular, according to the present invention, the examination for detecting and interpreting/analyzing the reaction product can be accurately and rapidly performed in one integrated system after the distribution of the sample, the reaction of the reagent with the sample, and the like, so that it is possible to shorten the examination time, improve the accuracy and reproducibility of the examination, and reduce the steps and input costs involved in the entire examination.

In addition, in the arm unit 550 provided in the automated liquid-phase immunofluorescence analysis apparatus 1 according to one embodiment of the present invention, the punch arm 552, the collection arm 556, and the pipette arm 554 are integrally provided and formed as an integrated module. Therefore, when the dispensing Pump (Pump dispenser) drives the punch, washes, and separates the dispensing tip 20 and the washing tip 30, the vertical position can be controlled by one driving motor. Therefore, unlike when each module is provided by its own drive motor, respectively, the size can be reduced and the manufacturing cost can be reduced. In addition, the arm units 550 are provided as an integrated module such that each arm is connected to one up-down driving part 544 and operates, but the arm units 550 are designed in a structure that does not interfere with each other when driven. Thus, by using the arm unit 550 provided as an integrated module, a great effect can be brought about in terms of reduction in the overall size of the apparatus and reduction in manufacturing cost.

In addition, the pump unit 506 included in the apparatus according to an embodiment of the present invention employs a motor capable of controlling a rotational micro-step, so that the amount thereof can be precisely adjusted when a sample, a reagent, or a reaction product is separated, dispensed, or discharged through the dispensing tip.

In addition, the apparatus according to one embodiment of the present invention has the detachment module 340, and thus the dispensing tip 20 and the wash tip 30, which are used end, can be easily separated from the dispenser module 500. In addition, since the above-described separation is performed by the detachment module 340, the dispensing tip 20 and the washing tip 30 may be reused after the separation.

In addition, the apparatus according to an embodiment of the present invention includes a standard block 360, and can perform examination by a ratio to the standard fluorescence using the standard fluorescence.

In summary, the apparatus according to an embodiment of the present invention is an automated immunodetector with convenience in which reagents are integrally prepared, and thus there is no need to separately prepare reagents, and for example, three different examinations can be performed simultaneously. In the case of the existing apparatus, only the same inspection can be performed at the same time. In addition, the apparatus according to one embodiment of the present invention employs an integrated module that can perform all steps of punching, dispensing, and washing reagents, and employs a system that can minimize the deviation of an optical system and an instrument using standard fluorescence. In addition, the reaction temperature of the reagents can be constantly controlled. In addition, since the dispensing tip and the washing tip, which are consumables, can be mounted on the cuvette again, a separate space (Trash) for discarding the tips is not required. However, in the case of instruments using consumable dispensing tips and wash tips, they are provided in a form in which the dispensing tip is discarded after it is used. This is because contamination cannot be used in the examination of other reagents. In addition, according to the apparatus of one embodiment of the present invention, the dispensing tip needs to be replaced in preparation for the reaction of other reagents during the reaction of the reagent in the first cuvette. At this time, the used tip is attached to the cuvette 1, and a preparation process is performed using the dispensing tips of the cuvettes 2, 3. Thereafter, the dispensing tip of the cuvette 1 is mounted again in preparation for a second incubation process. If the dispensing tip used at the first incubation is discarded, a new tip should be used in preparation for the second incubation. The instrument is characterized in that the tip is arranged on a cuvette, then after the other cuvettes are separately injected and mixed, the tip of the original cuvette is reused to carry out the next step of process, and therefore the consumption of the separately injected tip and the washing tip for consumption can be limited to one. In addition, since it is not necessary to prepare a dispensing tip inside the instrument, there is a space advantage and a more miniaturized apparatus can be designed.

Although the preferred embodiments of the present invention have been described in detail hereinabove, the scope of the right of the present invention is not limited thereto, and various modifications and improvements made by those skilled in the art using the basic concept of the present invention defined in the claims of the present invention also belong to the scope of the right of the present invention.