CN114609382A - A system for identifying immunoassays and methods of using the same - Google Patents

Abstract

The invention discloses a system for identifying immunoassay and an application method thereof, belonging to the technical field of immunoassay, comprising an immunoassay unit and a determination analysis unit, wherein the immunoassay unit is used for performing immunoassay detection process on a sample, the determination analysis unit is used for reading and analyzing a detection result obtained by the immunoassay unit based on an enzyme-linked immunoassay technology, and when in application, the system comprises the following specific steps: s1, preparing a reagent; s2, adding samples; s3, adding a substrate of the enzyme; s4, adding an enzyme reaction stopping solution; and S5, judging the result. The system for identifying immunoassay and the application method thereof avoid the difference of absorbance among micropores caused by different determination time in the determination process by adopting the optical analysis module in the vertical light path 8 or 12 channel detection mode, reflect the dose response curve of immunoassay by adopting a four-parameter equation and the like, perform auxiliary analysis on the result of qualitative determination, and reduce the probability of occurrence of false negative.

Description

A system for identifying immunoassays and methods of using the same

technical field

The invention belongs to the technical field of immunoassay, in particular to a system for identifying immunoassay and an application method thereof.

Background technique

Immunological detection is based on the principle of antigen-antibody specific reaction. Because it can use isotopes, enzymes, chemiluminescent substances, etc. to display or amplify the signal, it is often used to detect trace organisms such as proteins and hormones. active substance. Chemiluminescence immunoassay is a non-radioactive immunoassay technology that has developed rapidly in recent years. One of the important directions of detection.

In the prior art, when the concentration of the substance to be detected is high to a certain concentration, the phenomenon that the signal value is low due to the inability to form a double-antibody sandwich complex is called the high dose-hook effect (HD-HOOK effect). That is to say, the high-dose-hook effect refers to the high-dose segment of the dose-response curve in the double-site sandwich immunization assay. Only hooks, resulting in false negatives.

SUMMARY OF THE INVENTION

In view of the deficiencies of the prior art, the present invention provides a system for identifying immunoassays and an application method thereof, so as to solve the problems raised in the above background art.

In order to achieve the above purpose, the present invention provides the following technical solutions: a system for identifying immunoassays, comprising an immunoassay unit and an assay analysis unit, the immunoassay unit is used for performing an immunoassay detection process on a sample, and the assay The analysis unit is used to read and analyze the detection results obtained by the immune detection unit based on the enzyme-linked immunoassay detection technology, and determine the concentration of the antibody or antigen to be detected in the sample;

The immunoassay unit further includes a detection and analysis module, a sample tube identification module, a sample adding module and a reagent module, for performing immunoassay on the sample;

The measurement and analysis unit further includes an optical analysis module, a data analysis module and a result derivation module, which are used for qualitative and quantitative result judgment on the detection result.

Wherein, the optical analysis module in the assay analysis unit receives the sample from the immune detection unit, the output terminal of the optical analysis module is connected to the input terminal of the data analysis module, and the output terminal signal of the data analysis module is connected to the result The input of the export module.

To further optimize this technical solution, the reagent module of the immunoassay unit further includes a reagent kit, a reagent chamber and an incubation tray, the temperature of the reagent chamber is controlled at 20±3°C, and the temperature of the incubation tray is controlled at 37±0.5°C .

To further optimize the technical solution, the kit further includes immunosorbent, conjugate, enzyme substrate, enzyme-labeled working solution, conjugate and sample dilution, washing solution and enzyme reaction stop solution;

Further, the immunosorbent is a solid-phase carrier coated with an antigen or antibody, the conjugate is an antigen or antibody labeled with an enzyme, the substrate of the enzyme is tetramethylbenzidine TMB, and the enzyme The standard working solution was horseradish peroxidase HRP.

To further optimize this technical solution, the kit also includes a reference substance, a reference standard substance and a control serum, the reference substance is used in qualitative determination, and the reference substance includes a negative control substance and a positive control substance, and the reference substance Standards and control sera were used in quantitative assays.

To further optimize this technical solution, the sample adding module in the immunodetection unit further includes a sample needle sensing device, a sample needle flushing device and a diluent adding device, and the sample needle sensing device is used to sense the liquid level and sense the clot And the pressure induction of air bubbles, the sample needle flushing device uses a flushable sample probe, and cleans the inner wall and the outer wall at the same time, so that the carry rate is less than 0.1 ppm, and a disposable tip can also be used.

To further optimize the technical solution, the sample tube identification module in the immune detection unit is used to perform device loading operations on sample tubes of different specifications, and the sample tube identification module identifies the barcode on the sample tube.

The technical solution is further optimized. The optical analysis module in the measurement and analysis unit adopts the detection method of 8 or 12 channels of vertical optical paths. The measurement channel is a silicon light pipe or an optical fiber, and a reference channel is provided. Each measurement can be performed by itself. calibration.

To further optimize the technical solution, the data analysis module in the measurement and analysis unit is used to perform qualitative and quantitative measurement and analysis on the sample results, and the result export module is used to export the results of the measurement and analysis.

An application method for identifying an immunoassay is applied based on the above-mentioned system for identifying an immunoassay, comprising the following specific steps:

S1. Reagent preparation: Use the reagent module of the immunodetection unit to coat the solid-phase carrier with specific antibodies and incubate for a certain period of time to form solid-phase antibodies, and remove unbound antibodies and impurities after washing;

S2. Sample loading: The sample loading module of the immunodetection unit is used to incubate the antigen in the sample to fully react with the antibody on the solid-phase carrier to form a solid-phase antigen-antibody complex, and wash to remove other unbound substances;

S3. Add the substrate of the enzyme: the enzyme catalyzes the substrate on the chromogenic solid phase to produce a colored product through colorimetry to measure the amount of antigen in the sample;

S4. Add enzyme reaction termination solution: terminate the reaction of the sample in the detection and analysis module;

S5. Result determination: put the ELISA plate into the assay and analysis unit, and read and analyze the results through the assay and analysis unit to determine the qualitative and quantitative results.

To further optimize this technical solution, in S5, the qualitative and quantitative result determination further includes the following specific content:

1) During qualitative judgment, the positive judgment value and its gray area for determination are calculated through the calculation model. The gray area for measurement refers to a certain area where the measured absorbance is around the positive judgment value, and the result in this area is the qualitative judgment unqualified;

2) During the quantitative determination, the dose-response curve of the immunoassay can be reflected by the four-parameter equation, and the quantitative analysis of the immunoassay dose can also be performed through the connection point and linear regression calculation model.

Compared with the prior art, the present invention provides a system for identifying immunoassays and an application method thereof, which have the following beneficial effects:

The system for identifying immunoassays and the application method thereof, by adopting an optical analysis module in a vertical optical path 8 or 12-channel detection mode, avoids the difference in absorbance between microwells due to different measurement times during the measurement process, and at the same time , by quantitatively measuring the sample, using the four-parameter equation to reflect the dose-response curve of the immunoassay, and assisting the analysis of the qualitative judgment results to reduce the probability of false negatives.

Description of drawings

1 is a schematic structural diagram of a system for identifying immunoassays proposed by the present invention;

FIG. 2 is a schematic flow chart of an application method for identifying an immunoassay proposed by the present invention.

Detailed ways

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

Example:

Please refer to FIG. 1 , a system for identifying immunoassays, including an immunoassay unit and an assay analysis unit, the immunoassay unit is used for the detection process of immunoassay on a sample, and the assay assay unit is based on enzyme-linked immunoassay The technology is used to read and analyze the test results obtained by the immune detection unit, and determine the concentration of the antibody or antigen to be tested in the sample;

The immunoassay unit further includes a detection and analysis module, a sample tube identification module, a sample adding module and a reagent module, for performing immunoassay on the sample;

The measurement and analysis unit further includes an optical analysis module, a data analysis module and a result derivation module, which are used for qualitative and quantitative result judgment on the detection result.

Wherein, the optical analysis module in the assay analysis unit receives the sample from the immune detection unit, the output terminal of the optical analysis module is connected to the input terminal of the data analysis module, and the output terminal signal of the data analysis module is connected to the result The input of the export module.

Specifically, the reagent module of the immunoassay unit further includes a reagent kit, a reagent chamber and an incubation tray, the temperature of the reagent chamber is controlled at 20±3°C, and the temperature of the incubation tray is controlled at 37±0.5°C.

Specifically, the kit further includes an immunosorbent, a conjugate, an enzyme substrate, an enzyme-labeled working solution, a diluent of the conjugate and the sample, a washing solution, and an enzyme reaction stop solution;

Further, the immunosorbent is a solid-phase carrier coated with an antigen or antibody, the conjugate is an antigen or antibody labeled with an enzyme, the substrate of the enzyme is tetramethylbenzidine TMB, and the enzyme The standard working solution was horseradish peroxidase HRP.

Specifically, the kit further includes a reference substance, a reference standard substance and a control serum, the reference substance is used in qualitative determination, the reference substance includes a negative control substance and a positive control substance, the reference standard substance and Control sera were used in quantitative assays.

Specifically, the sample adding module in the immunoassay unit further includes a sample needle sensing device, a sample needle rinsing device and a diluent adding device, the sample needle sensing device is used for sensing the liquid level and sensing the level of clots and air bubbles. For pressure induction, the sample needle flushing device adopts a flushable sample probe, and simultaneously cleans the inner wall and the outer wall, so that the carry rate is less than 0.1 ppm, and a disposable tip can also be used.

Specifically, the sample tube identification module in the immune detection unit is used to perform device loading operations on sample tubes of different specifications, and the sample tube identification module identifies the barcode on the sample tube.

Specifically, the optical analysis module in the measurement and analysis unit adopts a detection method of 8 or 12 channels of vertical optical paths, the measurement channel is a silicon light pipe or an optical fiber, and a reference channel is provided, and self-calibration can be performed for each measurement. The difference in absorbance between microwells due to different measurement times during the measurement process is avoided by using an optical analysis module with 8 or 12-channel detection methods in a vertical optical path.

Specifically, the data analysis module in the measurement and analysis unit is used to perform qualitative and quantitative measurement and analysis on the sample results, and the result export module is used to export the result of the measurement and analysis.

Please refer to FIG. 2 , an application method for identifying an immunoassay is applied based on the above-mentioned system for identifying an immunoassay, comprising the following specific steps:

S1. Reagent preparation: Use the reagent module of the immunodetection unit to coat the solid-phase carrier with specific antibodies and incubate for a certain period of time to form solid-phase antibodies, and remove unbound antibodies and impurities after washing;

S2. Sample loading: The sample loading module of the immunodetection unit is used to incubate the antigen in the sample to fully react with the antibody on the solid-phase carrier to form a solid-phase antigen-antibody complex, and wash to remove other unbound substances;

S3. Add the substrate of the enzyme: the enzyme catalyzes the substrate on the chromogenic solid phase to produce a colored product through colorimetry to measure the amount of antigen in the sample;

S4. Add enzyme reaction termination solution: terminate the reaction of the sample in the detection and analysis module;

S5. Result determination: put the ELISA plate into the assay and analysis unit, and read and analyze the results through the assay and analysis unit to determine the qualitative and quantitative results.

By quantitatively measuring the samples, the four-parameter equation is used to reflect the dose-response curve of the immunoassay, and the qualitative judgment results are assisted by analysis to reduce the probability of false negatives.

Specifically, in the S5, the qualitative and quantitative result determination further includes the following specific content:

1) During qualitative judgment, the positive judgment value and its gray area for determination are calculated through the calculation model. The gray area for measurement refers to a certain area where the measured absorbance is around the positive judgment value, and the result in this area is the qualitative judgment unqualified;

2) During the quantitative determination, the dose-response curve of the immunoassay can be reflected by the four-parameter equation, and the quantitative analysis of the immunoassay dose can also be performed through the connection point and linear regression calculation model.

The beneficial effects of the present invention are: the system for identifying immunoassays and the application method thereof, by adopting the optical analysis module of the vertical optical path 8 or 12-channel detection mode, avoids the occurrence of micropores caused by different measurement times during the measurement process. At the same time, by quantitatively measuring the samples, using a four-parameter equation to reflect the dose-response curve of the immunoassay, and assisting in the analysis of the qualitative judgment results to reduce the probability of false negatives.

In the description of this specification, description with reference to the terms "one embodiment," "some embodiments," "example," "specific example," or "some examples", etc., mean specific features described in connection with the embodiment or example , structure, material or feature is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, those skilled in the art may combine and combine the different embodiments or examples described in this specification, as well as the features of the different embodiments or examples, without conflicting each other.

Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, and substitutions can be made in these embodiments without departing from the principle and spirit of the invention and modifications, the scope of the present invention is defined by the appended claims and their equivalents.

Claims (10)

1. A system for identifying an immunoassay, comprising an immunoassay unit and an assay analysis unit, wherein the immunoassay unit is used to perform an immunoassay detection process on a sample, and the assay assay unit is based on an enzyme-linked immunosorbent assay. The detection technology is used to read and analyze the detection results obtained by the immune detection unit to determine the concentration of the antibody or antigen to be detected in the sample; The immunoassay unit further includes a detection and analysis module, a sample tube identification module, a sample adding module and a reagent module, for performing immunoassay on the sample; The measurement and analysis unit further includes an optical analysis module, a data analysis module and a result derivation module, which are used for qualitative and quantitative result judgment on the detection result; Wherein, the optical analysis module in the assay analysis unit receives the sample from the immune detection unit, the output terminal of the optical analysis module is connected to the input terminal of the data analysis module, and the output terminal signal of the data analysis module is connected to the result The input of the export module. 2. A system for identifying immunoassays according to claim 1, wherein the reagent module of the immunoassay unit further comprises a reagent kit, a reagent compartment and an incubation tray, and the temperature of the reagent compartment is controlled at 20±3°C, and the temperature of the incubation plate was controlled at 37±0.5°C. 3. A system for identifying immunoassays according to claim 2, wherein the test kit further comprises an immunosorbent, a conjugate, an enzyme substrate, an enzyme-labeled working solution, a conjugate and a sample Diluent, washing solution and enzyme reaction stop solution; Further, the immunosorbent is a solid-phase carrier coated with an antigen or antibody, the conjugate is an antigen or antibody labeled with an enzyme, the substrate of the enzyme is tetramethylbenzidine TMB, and the enzyme The standard working solution was horseradish peroxidase HRP. 4. a kind of system for identifying immunoassay according to claim 3, is characterized in that, described test kit also includes reference substance, reference standard substance and control serum, and described reference substance is used in qualitative determination , the control substance includes negative control substance and positive control substance, and the reference standard substance and control serum are used in quantitative determination. 5 . The system for identifying immunoassays according to claim 1 , wherein the sample adding module in the immunoassay unit further comprises a sample needle sensing device, a sample needle rinsing device and a diluent adding device, 6 . The sample needle sensing device is used for sensing liquid level and pressure sensing of clots and air bubbles. Disposable tips are available. 6 . The system for identifying immunoassays according to claim 1 , wherein the sample tube identification module in the immunoassay unit is used to perform device loading operations on sample tubes of different specifications. The identification module identifies the barcode on the sample tube. 7. A system for identifying immunoassays according to claim 1, wherein the optical analysis module in the assay analysis unit adopts the detection mode of vertical optical path 8 or 12 channels, and the assay channel is a silicon light pipe or optical fiber, and is equipped with a reference channel, which can be self-calibrated for each measurement. 8. A system for identifying immunoassays according to claim 1, wherein the data analysis module in the assay analysis unit is used to perform qualitative and quantitative assay analysis on sample results, and the result export module Used to export the results of assay analysis. 9. a kind of application method for identifying immunoassay, apply based on a kind of system for identifying immunoassay described in claim 1-8, it is characterized in that, comprise the following concrete steps: S1. Reagent preparation: Use the reagent module of the immunodetection unit to coat the solid-phase carrier with specific antibodies and incubate for a certain period of time to form solid-phase antibodies, and remove unbound antibodies and impurities after washing; S2. Sample loading: The sample loading module of the immunodetection unit is used to incubate the antigen in the sample to fully react with the antibody on the solid-phase carrier to form a solid-phase antigen-antibody complex, and wash to remove other unbound substances; S3. Add the substrate of the enzyme: the enzyme catalyzes the substrate on the chromogenic solid phase to produce a colored product through colorimetry to measure the amount of antigen in the sample; S4. Add enzyme reaction termination solution: terminate the reaction of the sample in the detection and analysis module; S5. Result determination: put the ELISA plate into the assay and analysis unit, and read and analyze the results through the assay and analysis unit to determine the qualitative and quantitative results. 10. A kind of application method for identifying immunoassay according to claim 9, is characterized in that, in described S5, the qualitative and quantitative result judgment further comprises the following specific content: 1) During qualitative judgment, the positive judgment value and its gray area for determination are calculated through the calculation model. The gray area for measurement refers to a certain area where the measured absorbance is around the positive judgment value, and the result in this area is the qualitative judgment unqualified; 2) During the quantitative determination, the dose-response curve of the immunoassay can be reflected by the four-parameter equation, and the quantitative analysis of the immunoassay dose can also be performed through the connection point and linear regression calculation model.

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