Enzyme-linked immunosorbent assay kit for quantitatively detecting human-serum thyrotrophin receptor antibodies and detecting method

An enzyme-linked immunoassay and thyrotropin technology, which is applied in the field of immunoassays, can solve the problems of low positive detection rate, high price and the like, and achieves the effects of low price, simple method, and easy popularization and application.

Inactive Publication Date: 2012-10-17
GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The present invention aims to solve the problem that the positive detection rate of human serum TRAb, especially TSBAb, is low or the price is very expensive, and provides a quantitative

Method used

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  • Enzyme-linked immunosorbent assay kit for quantitatively detecting human-serum thyrotrophin receptor antibodies and detecting method
  • Enzyme-linked immunosorbent assay kit for quantitatively detecting human-serum thyrotrophin receptor antibodies and detecting method
  • Enzyme-linked immunosorbent assay kit for quantitatively detecting human-serum thyrotrophin receptor antibodies and detecting method

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Embodiment Construction

[0031] The present invention will be described in detail below in conjunction with the accompanying drawings and embodiments.

[0032] Main reagents and raw materials:

[0033] 1. Fusion protein antigen of the carboxy-terminal fragment of the extramembrane region of recombinant hTSHR (prepared by ourselves)

[0034] 2. Carbonate buffer (prepared by yourself)

[0035] 3. Phosphate rinse solution (PBST) (prepared by yourself)

[0036] 4. Blocking solution (prepared by yourself)

[0037] 5. Thyrotropin receptor antibody standard solution (prepared by ourselves)

[0038] 6. Sample diluent (prepared by yourself)

[0039] 7. HRP-labeled goat anti-human IgG secondary antibody (commercially available)

[0040] 8. Chromogenic substrate Tetramethylbenzidine (TMB) (commercially available)

[0041] 9. Reaction termination solution 2M H 2 SO 4 (self-prepared)

[0042] 10. Quality control serum (prepared by yourself)

[0043] 1. Preparation of the fusion protein (TrxFus-hTSHRc) anti...

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Abstract

The invention discloses an enzyme-linked immunosorbent assay kit for quantitatively detecting a human-serum thyrotrophin receptor antibody and a detecting method. The kit contains a 96 ferment plate precoated with segment fusion protein at the carboxyl terminal of the extracellular domain of a recombinant human thyroid stimulating hormone receptor and the following bottled reagents: phosphate bleaching liquid, confining liquid, sample diluents, thyrotrophin receptor antibody standard liquid, goat-anti-human IgG second antibody labeled with horse radish peroxidase, chromogenic substrate (tetramethyl benzidine), reaction stopping solution and quality-control serum. The invention has the advantages that the detected human-serum thyrotrophin receptor antibodies give priority to a thyroid stimulation blocking antibody, the positive detection rate to patients with hashimoto thyroiditis is high, the detected result is accurate and objective, the method is simple and convenient, and the promotion and the application are easy. The invention has an important significance in diagnosis, guide therapy and curative effect evaluation of the autoimmune thyroid disorders, and has an important clinic reference value in plan making and method selection for treating Graves disease.

Description

technical field [0001] The invention belongs to the immunoassay method, specifically, a solid-phase antigen is prepared by using a porous microplate as a carrier for quantitative detection of human serum thyrotropin receptor antibody (TRAb), especially thyroid stimulating blocking antibody (TSBAb) )-based enzyme-linked immunoassay (ELISA) kits and detection methods. Background technique [0002] Autoimmune thyroid disease (AITD) is a group of organ-specific autoimmune diseases with complex pathogenesis, including Graves' disease (GD), Hashimoto's thyroiditis (HT), etc. Human thyroid-stimulating hormone receptor (hTSHR) is the main target antigen of AITD and plays an important role in the pathogenesis of AITD. hTSHR is a glycoprotein present on the membrane of human thyroid follicular epithelial cells (TEC). After it combines with thyrotropin (TSH), it regulates the growth, differentiation, synthesis and release of thyroid hormones of TEC. On the basis of environmental and ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
Inventor 李宁方佩华陈慧谭建柴锦燕
Owner GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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