CN114609382A - System for identifying immunoassay and application method thereof - Google Patents
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- CN114609382A CN114609382A CN202210266732.XA CN202210266732A CN114609382A CN 114609382 A CN114609382 A CN 114609382A CN 202210266732 A CN202210266732 A CN 202210266732A CN 114609382 A CN114609382 A CN 114609382A
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- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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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
Technical Field
The invention belongs to the technical field of immunoassay, and particularly relates to a system for identifying immunoassay and an application method thereof.
Background
Immunological detection is based on the principle of antigen-antibody specific reaction, and is often used for detecting a trace amount of bioactive substances such as proteins and hormones because it allows display of a sample or amplification of a signal using an isotope, enzyme, chemiluminescent substance, or the like. Chemiluminescence immunoassay is a non-radioactive immunoassay which is developed rapidly in recent years, and the principle is that a chemiluminescence substance is used for amplifying signals and an immunological binding process is directly measured by virtue of the luminous intensity, and the method is one of important directions of immunological detection.
In the prior art, when the concentration of a substance to be detected is high to a certain concentration, a phenomenon that a double-antibody sandwich complex cannot be formed so that a signal value is low is called a high dose-HOOK effect (HD-HOOK effect). That is, the high dose-hook effect refers to the phenomenon that in the double-site sandwich immunoassay, the linear trend of the high dose section of the dose response curve is not in a platform shape and extends backwards infinitely, but is curved downwards to be similar to a hook, so that false negative is generated.
Disclosure of Invention
In view of the deficiencies of the prior art, the present invention provides a system for identification of immunoassays and a method for using the same to solve the problems set forth in the background art described above.
In order to achieve the purpose, the invention provides the following technical scheme: a system for identifying immunoassay comprises an immunoassay unit and a determination analysis unit, wherein the immunoassay unit is used for performing immunoassay detection process on a sample, and 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 judging the concentration of an antibody or an antigen to be detected in the sample;
the immunoassay unit further comprises a detection analysis module, a sample tube identification module, a sample adding module and a reagent module, and is used for performing immunoassay on a sample;
the determination analysis unit further comprises an optical analysis module, a data analysis module and a result derivation module, and is used for carrying out qualitative and quantitative result judgment on the detection result.
The optical analysis module in the determination analysis unit receives a sample from the immunodetection unit, the output end of the optical analysis module is connected with the input end of the data analysis module in a signal mode, and the output end of the data analysis module is connected with the input end of the result derivation module in a signal mode.
Further optimizing the technical scheme, the reagent module of the immunity detection unit further comprises a reagent kit, a reagent cabin and an incubation disc, wherein the temperature of the reagent cabin is controlled to be 20 +/-3 ℃, and the temperature of the incubation disc is controlled to be 37 +/-0.5 ℃.
Further optimizing the technical scheme, the kit further comprises an immunoadsorbent, a conjugate, a substrate of enzyme, an enzyme-labeled working solution, a diluent of the conjugate and a sample, a washing solution and an enzyme reaction stopping solution;
furthermore, the immunoadsorbent is a solid phase carrier coated with antigen or antibody, the conjugate is enzyme-labeled antigen or antibody, the substrate of the enzyme is tetramethylbenzidine TMB, and the enzyme-labeled working solution is horseradish peroxidase HRP.
The technical scheme is further optimized, the kit further comprises a reference substance, a reference standard substance and control serum, the reference substance is used in qualitative determination, the reference substance comprises a negative reference substance and a positive reference substance, and the reference standard substance and the control serum are used in quantitative determination.
Further optimizing the technical scheme, the sample adding module in the immunoassay unit further comprises a sample needle sensing device, a sample needle flushing device and a diluent adding device, wherein the sample needle sensing device is used for sensing the liquid level and sensing the pressure sensing of clots and bubbles, the sample needle flushing device adopts a sample probe capable of being flushed, and simultaneously cleans the inner wall and the outer wall, so that the carrying rate is less than 0.1ppm, and a disposable suction head can also be adopted.
Further optimizing the technical scheme, the sample tube identification module in the immunodetection unit is used for carrying out equipment sample loading operation on sample tubes with different specifications, and the sample tube identification module identifies the bar codes on the sample tubes.
Further optimizing the technical scheme, the optical analysis module in the measurement and analysis unit adopts a detection mode of a vertical light path 8 or 12 channel, the measurement channel is a silicon light tube or an optical fiber, and is provided with a reference channel, and self-calibration can be carried out in each measurement.
Further optimizing the technical scheme, the data analysis module in the measurement analysis unit is used for performing qualitative and quantitative measurement analysis on the sample result, and the result derivation module is used for deriving the result after the measurement analysis.
An application method for identifying immunoassay based on the system for identifying immunoassay comprises the following specific steps:
s1, preparation of reagents: incubating a solid phase carrier coated with a specific antibody for a certain time by using a reagent module of an immunoassay unit to form a solid phase antibody, and washing to remove unbound antibody and impurities;
s2, sample adding: incubating by adopting a sample adding module of the immunoassay unit to ensure that the antigen in the sample fully reacts with the antibody on the solid phase carrier to form a solid phase antigen-antibody complex, and washing to remove other unbound substances;
s3, substrate to add enzyme: the enzyme on the chromogenic solid phase catalyzes a substrate to generate a colored product, and the amount of the antigen in the sample is measured through colorimetry;
s4, adding an enzyme reaction stopping solution: terminating the reaction of the sample in the detection and analysis module;
s5, judging the result: the microplate is placed in a measurement and analysis unit, and the measurement and analysis unit reads and analyzes the result to determine the qualitative and quantitative result.
Further optimizing the technical solution, in S5, the determining of the qualitative and quantitative result further includes the following specific contents:
1) when qualitative judgment is carried out, a positive judgment value and a gray measurement area thereof are calculated through a calculation model, wherein the gray measurement area refers to a certain area with the measurement absorbance around the positive judgment value, and the result in the area is unqualified for qualitative judgment;
2) when quantitative determination is carried out, a four-parameter equation is used for reflecting a dose response curve of immunoassay, and quantitative analysis of immunoassay dose can also be carried out through a connected point and linear regression calculation model.
Compared with the prior art, the invention provides a system for identifying immunoassay and an application method thereof, and the system has the following beneficial effects:
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 a vertical light path 8 or 12 channel detection mode, and simultaneously, the quantitative determination is carried out on a sample, a four-parameter equation and the like are adopted to reflect the dose response curve of immunoassay, the result of qualitative determination is subjected to auxiliary analysis, and the probability of false negative is reduced.
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FIG. 1 is a schematic diagram of a system for identifying immunoassays according to the present invention;
FIG. 2 is a schematic flow diagram of an application method for an identification immunoassay according to the present invention.
Detailed Description
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, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example (b):
referring to fig. 1, a system for identifying an immunoassay includes an immunoassay unit and a measurement and analysis unit, wherein the immunoassay unit is used for performing a detection process of immunoassay on a sample, and the measurement and analysis unit is used for reading and analyzing a detection result obtained by the immunoassay unit based on an enzyme-linked immunoassay technology, and determining the concentration of an antibody or an antigen to be detected in the sample;
the immunoassay unit further comprises a detection analysis module, a sample tube identification module, a sample adding module and a reagent module, and is used for performing immunoassay on a sample;
the determination analysis unit further comprises an optical analysis module, a data analysis module and a result derivation module, and is used for carrying out qualitative and quantitative result judgment on the detection result.
The optical analysis module in the determination analysis unit receives a sample from the immunodetection unit, the output end of the optical analysis module is connected with the input end of the data analysis module in a signal mode, and the output end of the data analysis module is connected with the input end of the result derivation module in a signal mode.
Specifically, the reagent module of the immunoassay unit further comprises a reagent kit, a reagent cabin and an incubation disc, wherein the temperature of the reagent cabin is controlled to be 20 +/-3 ℃, and the temperature of the incubation disc is controlled to be 37 +/-0.5 ℃.
Specifically, the kit further comprises an immunoadsorbent, a conjugate, an enzyme substrate, an enzyme-labeled working solution, a diluent of the conjugate and a sample, a washing solution and an enzyme reaction stopping solution;
furthermore, the immunoadsorbent is a solid phase carrier coated with antigen or antibody, the conjugate is enzyme-labeled antigen or antibody, the substrate of the enzyme is tetramethylbenzidine TMB, and the enzyme-labeled working solution is horseradish peroxidase HRP.
Specifically, the kit further comprises a reference substance, a reference standard substance and control serum, wherein the reference substance is used in qualitative determination, the reference substance comprises a negative reference substance and a positive reference substance, and the reference standard substance and the control serum are used in quantitative determination.
Specifically, the sample adding module in the immunoassay unit further comprises a sample needle sensing device, a sample needle flushing device and a diluent adding device, wherein the sample needle sensing device is used for sensing the liquid level and sensing the pressure sensing of clots and bubbles, the sample needle flushing device adopts a flushable sample probe and simultaneously cleans the inner wall and the outer wall, the carrying rate is less than 0.1ppm, and a disposable suction head can also be adopted.
Specifically, a sample tube identification module in the immunoassay unit is used for performing device sample loading operation on sample tubes of different specifications, and the sample tube identification module identifies a barcode on the sample tube.
Specifically, the optical analysis module in the measurement and analysis unit adopts a detection mode of a vertical light path 8 or 12 channel, the measurement channel is a silicon light tube or an optical fiber, and is provided with a reference channel, and self-calibration can be carried out in each measurement. The optical analysis module adopting the vertical light path 8 or 12 channel detection mode avoids the difference of absorbance among the micropores caused by different measurement time in the measurement process.
Specifically, the data analysis module in the measurement analysis unit is configured to perform qualitative and quantitative measurement analysis on the sample result, and the result derivation module is configured to derive the result after the measurement analysis.
Referring to FIG. 2, a method for applying an identification immunoassay based on the above-mentioned system for identifying an immunoassay comprises the following steps:
s1, preparation of reagents: incubating the specific antibody coated solid phase carrier for a certain time by adopting a reagent module of the immunoassay unit to form a solid phase antibody, and removing unbound antibody and impurities after washing;
s2, sample adding: incubating by adopting a sample adding module of the immunoassay unit to ensure that the antigen in the sample fully reacts with the antibody on the solid phase carrier to form a solid phase antigen-antibody complex, and washing to remove other unbound substances;
s3, substrate to add enzyme: the enzyme on the chromogenic solid phase catalyzes a substrate to generate a colored product, and the amount of the antigen in the sample is measured through colorimetry;
s4, adding an enzyme reaction stopping solution: terminating the reaction of the sample in the detection and analysis module;
s5, judging the result: the microplate is placed in a measurement and analysis unit, and the measurement and analysis unit reads and analyzes the result to determine the qualitative and quantitative result.
The quantitative determination is carried out on the sample, a four-parameter equation and the like are adopted to reflect the dose response curve of immunoassay, the auxiliary analysis is carried out on the result of qualitative determination, and the probability of false negative occurrence is reduced.
Specifically, in S5, the qualitative and quantitative result determination further includes the following specific contents:
1) when qualitative judgment is carried out, a positive judgment value and a gray measurement area thereof are calculated through a calculation model, wherein the gray measurement area refers to a certain area with the measurement absorbance around the positive judgment value, and the result in the area is unqualified for qualitative judgment;
2) when quantitative determination is carried out, a four-parameter equation is used for reflecting a dose response curve of immunoassay, and quantitative analysis of immunoassay dose can also be carried out through a connected point and linear regression calculation model.
The invention has the beneficial effects that: 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 a vertical light path 8 or 12 channel detection mode, and simultaneously, the quantitative determination is carried out on a sample, a four-parameter equation and the like are adopted to reflect the dose response curve of immunoassay, the result of qualitative determination is subjected to auxiliary analysis, and the probability of false negative is reduced.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer 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, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. The system for identifying immunoassay is characterized by comprising an immunoassay unit and a determination analysis unit, wherein the immunoassay unit is used for performing immunoassay detection process on a sample, and 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 judging the concentration of an antibody or an antigen to be detected in the sample;
the immunoassay unit further comprises a detection analysis module, a sample tube identification module, a sample adding module and a reagent module, and is used for performing immunoassay on a sample;
the measuring and analyzing unit further comprises an optical analyzing module, a data analyzing module and a result deriving module, and is used for carrying out qualitative and quantitative result judgment on the detection result;
the optical analysis module in the determination analysis unit receives a sample from the immunodetection unit, the output end of the optical analysis module is connected with the input end of the data analysis module in a signal mode, and the output end of the data analysis module is connected with the input end of the result derivation module in a signal mode.
2. The system for identifying an immunoassay according to claim 1, wherein the reagent module of the immunoassay unit further comprises a reagent kit, a reagent compartment and an incubation tray, the temperature of the reagent compartment is controlled at 20 ± 3 ℃ and the temperature of the incubation tray is controlled at 37 ± 0.5 ℃.
3. The system for identifying an immunoassay according to claim 2, wherein the kit further comprises an immunoadsorbent, a conjugate, a substrate for an enzyme, an enzyme-labeled working solution, a diluent for the conjugate and the sample, a washing solution, and an enzyme reaction termination solution;
furthermore, the immunoadsorbent is a solid phase carrier coated with antigen or antibody, the conjugate is enzyme-labeled antigen or antibody, the substrate of the enzyme is tetramethylbenzidine TMB, and the enzyme-labeled working solution is horseradish peroxidase HRP.
4. The system of claim 3, wherein the kit further comprises a control, a reference standard and a control serum, wherein the control is used in a qualitative assay, the control comprises a negative control and a positive control, and the reference standard and the control serum are used in a quantitative assay.
5. The system of claim 1, wherein the sample application module of the immunoassay unit further comprises a sample needle sensing device for sensing a liquid level and sensing pressure sensing of clots and air bubbles, a sample needle washing device for washing the sample probe while washing the inner and outer walls so that the carrying rate is less than 0.1ppm, and a disposable tip.
6. The system of claim 1, wherein the sample tube identification module of the immunoassay unit is configured to perform a device loading operation on sample tubes of different sizes, and the sample tube identification module identifies a barcode on the sample tube.
7. The system of claim 1, wherein the optical analysis module of the assay analysis unit adopts a detection mode of vertical light path 8 or 12 channels, the measurement channel is a silicon light tube or an optical fiber, and is provided with a reference channel, and each measurement can be self-calibrated.
8. The system of claim 1, wherein the data analysis module of the assay analysis unit is configured to perform qualitative and quantitative assay analysis on the sample results, and the result derivation module is configured to derive the results of the assay analysis.
9. A method for the use of an identification immunoassay based on a system for the identification immunoassay according to claims 1 to 8, comprising the following specific steps:
s1, preparation of reagents: incubating the specific antibody coated solid phase carrier for a certain time by adopting a reagent module of the immunoassay unit to form a solid phase antibody, and removing unbound antibody and impurities after washing;
s2, sample adding: incubating by adopting a sample adding module of the immunoassay unit to ensure that the antigen in the sample fully reacts with the antibody on the solid phase carrier to form a solid phase antigen-antibody complex, and washing to remove other unbound substances;
s3, substrate to add enzyme: the enzyme on the chromogenic solid phase catalyzes a substrate to generate a colored product, and the amount of the antigen in the sample is measured through colorimetry;
s4, adding an enzyme reaction stopping solution: terminating the reaction of the sample in the detection and analysis module;
s5, judging the result: the microplate is placed in a measurement and analysis unit, and the measurement and analysis unit reads and analyzes the result to determine the qualitative and quantitative result.
10. The method of claim 9, wherein the step of determining the qualitative or quantitative result in S5 further comprises the following steps:
1) when qualitative judgment is carried out, a positive judgment value and a gray measurement area thereof are calculated through a calculation model, wherein the gray measurement area refers to a certain area with the measurement absorbance around the positive judgment value, and the result in the area is unqualified for qualitative judgment;
2) when quantitative determination is carried out, a four-parameter equation is used for reflecting a dose response curve of immunoassay, and quantitative analysis of immunoassay dose can also be carried out through a connected point and linear regression calculation model.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114778867A (en) * | 2022-06-13 | 2022-07-22 | 深圳市帝迈生物技术有限公司 | Sample detection device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114778867A (en) * | 2022-06-13 | 2022-07-22 | 深圳市帝迈生物技术有限公司 | Sample detection device |
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