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Method for quantitatively detecting TRAb plate-type fluorescent enzyme immunity and application thereof

A technology for quantitative detection and human serum, which is applied in the field of medical ultra-trace detection, can solve the problems of unsuitable clinical routine detection and immature technology, and achieve the effects of easy clinical promotion and application, low price and convenient operation

Inactive Publication Date: 2010-04-07
GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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  • Abstract
  • Description
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Problems solved by technology

This method can distinguish TSAb and TSBAb, but it is still a preliminary study, the technology is not yet mature, and it cannot be used in clinical routine detection

Method used

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  • Method for quantitatively detecting TRAb plate-type fluorescent enzyme immunity and application thereof
  • Method for quantitatively detecting TRAb plate-type fluorescent enzyme immunity and application thereof
  • Method for quantitatively detecting TRAb plate-type fluorescent enzyme immunity and application thereof

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Embodiment Construction

[0018] Plate-type solid-phase quantitative detection of human TRAb (mainly TSAb) FEIA of the present invention uses recombinant hTSHR extramembrane amino-terminal protein as an antigen-coated porous plate, and seals it with PBST containing 1-5% BSA; then uses the porous plate as a carrier Add anti-hTSHR primary antibody, enzyme-labeled secondary antibody, fluorescent substrate and stop solution in sequence, and finally measure the fluorescence intensity on a fluorescence luminescence analyzer.

[0019] The amino acid sequence of the amino-terminal protein of the recombinant human thyroid-stimulating hormone receptor hTSHR extramembrane region of the antigen is:

[0020] Met Gly Cys Ser Ser Pro Pro Cys Glu Cys His Gln Glu Glu Asp Phe 16AA

[0021] 1 7 13

[0022] Arg Val Thr Cys lys Asp Ile Gln Arg Ile Pro Ser Leu Pro Pro Ser 32AA

[0023] 19 25 31

[0024] Thr Gln Thr Leu lys Leu Ile Glu Thr His Leu Arg Thr Ile Pro Ser 48AA

[0025] 37 43

[0026]...

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Abstract

The invention relates to a method for quantitatively detecting TRAb plate-type fluorescent enzyme immunity and an application thereof. The method is characterized in that recombination human thyroxin stimulating hormone receptor (hTSHR) extracellular domain amino terminal proteins is taken as antigen to coat a perforated plate and then is sealed by PBST containing 1-5% of BSA; anti-hTSHR first antibody or standard substance, enzyme-labeled second antibody, fluorescent substrate and stopping solution are sequentially added by utilizing the perforated plate as a carrier; and finally fluorescence intensity is measured on a fluorescence illuminant analyzer. The clinical detection of TRAb which mainly comprises TSAb in patient serum has great significance on pathogenic diagnosis, curative effect observation and drug withdrawal occasion determination of GD hyperthyroidism. The invention performs the fluorescence enzyme immunoassay by utilizing a multi-hole site plastic plate as a solid carrier, has easy sheet material source, low cost and easy promotion, and the method is distinctive, sensitive, accurate and stable, thereby being suitable for clinical requirement and promotion.

Description

【Technical field】 [0001] The invention belongs to the technical field of medical ultra-trace detection, and uses a porous plate as a carrier to prepare a solid-phase antigen microplate plate for quantitative detection of human serum TRAb (mainly TSAb) plate-type fluorescent enzyme immunoassay (FEIA) and its application. 【Background technique】 [0002] Graves' disease (GD) is a multiple autoimmune thyroid disease (AITD), and thyrotropin receptor (TSHR) is the most important autoantigen in the pathogenesis of GD. TSHR is a glycoprotein present on the membrane of 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 genetic factors, abnormal changes in the body's immune function lead to AITD. TSHR is one of the main autoantigens of AITD, which stimulates the body to produce thyrotropin receptor antibody (TRAb), which is a h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/64G01N33/564C07K14/72
Inventor 方佩华李宁陈慧王少艳谭建
Owner GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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