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Thyroid stimulating hormone receptor antigen reagent and thyroid stimulating hormone receptor antibody quantitative detection kit

A thyroid stimulating hormone and kit technology, which can be used in biological tests, measuring devices, material inspection products, etc., and can solve problems such as interference and complex labeling processes.

Pending Publication Date: 2022-03-01
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are other commercial kits on the market that use the principle of the competition method to detect TRAb, and their detection systems can be roughly divided into two types: biotin-avidin system or FITC-anti-FITC system, both of which have certain drawbacks : Biotin-avidin system: When a certain amount of biotin in the specimen will interfere with the test results (reference 3: Zhang Guofeng, Guo Rui, Guan Haixia. Pay attention to information other than the test sheet - it is detected by biotin interference test An example of misdiagnosed as Graves' disease hyperthyroidism discusses the elements of diagnosing thyroid diseases [J]. Chinese Journal of Endocrinology and Metabolism, 2017, 33(09):723-725.); FITC-anti-FITC system: Some undergoing fluorescein angiography Intravenous fluorescein interference with clinical laboratory tests.Am JOphthalmol.1989Oct 15;108(4):375-9.doi:10.1016 / s0002-9394(14)73304-5.PMID:2801858.)
Both of the above systems involve a complex marking process and introduce corresponding interference factors

Method used

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  • Thyroid stimulating hormone receptor antigen reagent and thyroid stimulating hormone receptor antibody quantitative detection kit
  • Thyroid stimulating hormone receptor antigen reagent and thyroid stimulating hormone receptor antibody quantitative detection kit
  • Thyroid stimulating hormone receptor antigen reagent and thyroid stimulating hormone receptor antibody quantitative detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] A human thyroid-stimulating hormone receptor antibody quantitative detection kit, comprising: magnetic particles coated with anti-human TSHR mouse monoclonal antibody A, antigen reagent (anti-human TSHR mouse monoclonal antibody B-recombinant human TSHR complex), Sample diluent, horseradish peroxidase-labeled human stimulating thyrotropin receptor antibody and stimulating thyrotropin receptor antibody serial calibrator, the preparation process is as follows:

[0058] (1) Preparation of magnetic particle suspension coated with anti-human TSHR mouse monoclonal antibody A:

[0059] The carboxyl magnetic microspheres were activated under acidic conditions with EDC and NHS, the activation buffer was 0.1M MES (2-(N-morpholino)ethanesulfonic acid) buffer, and the activation time was 30min. After the activation was completed, Under the action of a magnetic field, the carboxyl magnetic microspheres are separated from the liquid, the supernatant is discarded, and an appropriate a...

Embodiment 2

[0078] Embodiment 2 stability evaluation

[0079] Stability evaluation method: using the kits provided in Comparative Example 1 and Example 1 to detect in parallel a series of stimulating thyrotropin receptor antibody concentrations 0IU / L, 2IU / L, 5IU / Calibrator of L, 10IU / L, 20IU / L, 50IU / L is measured on the automatic chemiluminescence instrument A2000 Plus system according to the detection steps, and the luminescence value of each group is obtained, and the signal value drop ≤ 10% is regarded as stable Good performance, the results are shown in Table 1, figure 2 , image 3 and Figure 4 , Table 1 is the stability evaluation result of the kit that the embodiment of the present invention and comparative example provide, figure 2 It is the stability evaluation result of the antigen reagent provided in the examples and comparative examples of the present invention at 20-25°C in the lyophilized solution, wherein curve a is the stability evaluation result of the antigen reage...

Embodiment 3

[0085] Embodiment 3 kit comparison

[0086] Quantitative detection kit for human thyroid stimulating hormone receptor antibody constructed according to Example 1 and kit C: quantitative detection kit for thyroid stimulating hormone receptor antibody (electrochemiluminescence method), kit D: human thyroid stimulating hormone receptor antibody The detection kit (enzyme-catalyzed chemiluminescence method) is used for detection and comparison of clinical samples, and the repeatability and interference of these three detection and quantification kits are analyzed and compared at the same time.

[0087] The detection principle of kit C is: competition method, magnetic particles coated with streptavidin, human immunocomplexes labeled with soluble porcine TSHR and biotinylated anti-porcine TSHR C-terminal mouse monoclonal antibody and ruthenium complex Monoclonal autoantibody M22 was tested;

[0088] The detection principle of kit D is: competition method, using FITC antibody to coat...

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Abstract

The invention provides a human thyroid stimulating hormone receptor antibody quantitative detection kit. The human thyroid stimulating hormone receptor antibody quantitative detection kit comprises a solid phase carrier coated with an anti-TSHR monoclonal antibody A; an antigen reagent, wherein the antigen reagent is an immune complex of an anti-TSHR monoclonal antibody B and a TSHR; a human stimulated thyroid stimulating hormone receptor antibody marked by a marker is contained; the anti-TSHR monoclonal antibody A and the anti-TSHR monoclonal antibody B do not generate competitive binding. The kit provided by the invention adopts a competition method reaction principle to detect the concentration of the thyroid stimulating hormone receptor antibody in serum or plasma, the kit is relatively stable, the interference of biotin or fluorescein sodium can be avoided, the preparation process is simpler and more stable, the repeatability is good, and the cost is greatly reduced while the product performance is improved.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a thyroid-stimulating hormone receptor antigen reagent and a thyroid-stimulating hormone receptor antibody quantitative detection kit. Background technique [0002] Thyroid-stimulating hormone receptor (TSHR) is a macromolecular glycoprotein located on the membrane of thyroid follicular epithelial cells and belongs to the G protein-coupled receptor family. Under physiological conditions, thyroid-stimulating hormone (TSH) binds to its receptor TSHR, and is activated by G protein to generate a cascade reaction to achieve biological effects. Under the influence of heredity and environment, the structure of TSHR changes, resulting in antigenicity, causing autoimmune reactions, and producing a large number of thyroid-stimulating hormone receptor antibodies (TRAb). TRAb is a group of heterogeneous antibodies, which can be divided into thyroid stimulating antibody (TSAb), th...

Claims

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Application Information

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IPC IPC(8): G01N33/76G01N33/577G01N33/543G01N33/58
CPCG01N33/76G01N33/577G01N33/54326G01N33/54393G01N33/581
Inventor 徐海娈靳增明余春霞赵艳芳韩阳瑞田晓平李彬
Owner AUTOBIO DIAGNOSTICS CO LTD
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