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Kit and method for detection of thyroid peroxidase antibody

A technology of peroxidase and thyroid, which is applied in the direction of chemiluminescence/bioluminescence, analysis by causing chemical reactions of materials, and measurement devices, etc. It can solve the problems of narrow detection range, long detection time of kits, low sensitivity of kits, etc. problems, to achieve the effect of wide detection range, improved environmental protection performance, simple operation and time saving

Inactive Publication Date: 2016-04-06
SUZHOU HAOOUBO BIOPHARML
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Prior art, such as publication number is CN1595161A, the patent of invention that disclosure date is 2005-3-16 discloses a kind of thyroid peroxidase antibody magnetic separation enzyme-linked immunoassay detection method, but the sensitivity of the test kit in this patent is still relatively high. Low (5IU / ml), and the detection time of the kit is too long (>90min); Another example is the invention patent with the publication number CN103336115A and the publication date of 2013-10-02 disclosing a thyroid peroxidase antibody magnetic separation enzyme However, the sensitivity of the kit in this patent is still low (1IU / ml), the detection time of the kit is too long (>90min), and the detection range is narrow (1-400IU / ml)

Method used

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  • Kit and method for detection of thyroid peroxidase antibody
  • Kit and method for detection of thyroid peroxidase antibody
  • Kit and method for detection of thyroid peroxidase antibody

Examples

Experimental program
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Effect test

Embodiment 1

[0038] The invention provides a kit for detecting thyroid peroxidase antibody and the preparation of matching reagents.

[0039] (1) Preparation of the first reagent:

[0040] Materials and instruments:

[0041] 1. Thyroid peroxidase antigen, prepared by HytestLTD, with a purity of 98%, stored in phosphate buffer;

[0042] 2. The coupling agent N-hydroxysuccinimide biotin ester (BNHS) was purchased from THERMO, and the reagents such as dimethyl sulfoxide (DMSO) and PBS were chemically pure;

[0043] 3. Analyze the balance.

[0044] Steps:

[0045] 1. Dialysis the thyroid peroxidase antigen with 0.02mol / L, pH7.4 PBS buffer at 4°C overnight, and change the medium once;

[0046] 2. Equilibrate the BNHS to room temperature (20°C), weigh 1.5mg of BNHS in 326μL of DMSO with a 1 / 10,000 analytical balance (maximum weighing 200g), and prepare 10mM BNHS (prepared for use now);

[0047] 3. According to the molar ratio of thyroid peroxidase antigen and BNHS of 1:20, add the solution obtained in step 2 ...

Embodiment 2

[0086] Example 2: Implementation of testing and evaluation of testing results

[0087] Materials and instruments:

[0088] 1. The kit prepared in Example 1;

[0089] 2. The luminescent substrate and cleaning fluid are produced by Suzhou Hao Ober Biomedical Co., Ltd.

[0090] 3. Chemiluminescence analyzer: BHP9507 chemiluminescence immunoassay analyzer produced by Beijing Hamamatsu Photonics Co., Ltd. or Lumiray1200, Lumiray1220, Lumiray1230, Lumiray1260, Lumiray1280 automatic chemiluminescence analysis and determination produced by Shenzhen Leidu Life Science Co., Ltd. instrument.

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Abstract

The present invention discloses a kit and method for detection of thyroid peroxidase antibody. The kit includes thyroid peroxidase antibody series standards, a magnetic separation reagent of a magnetic particle suspension conjugated with streptavidin and pigment, a first reagent of an antigen solution containing biotin N-hydroxysuccinimide ester labeled thyroid peroxidase antigen, a second reagent of a solution containing alkaline phosphatase labeled mouse-anti-human IgG monoclonal antibody. The kit is used for detection of thyroid peroxidase antibody. By the above-described manner, the invention uses a biotin-streptavidin signal amplification system and alkaline phosphatase labeling to solve the traditional problem of low ELISA sensitivity; the nanometer magnetic particle separation system achieves high sensitivity for radioimmunoassay (RIA), but has no radioactive contamination, and has greatly improved period of validity, security and environmental performance.

Description

Technical field [0001] The present invention relates to the technical field of in vitro diagnostics, in particular to a kit and method for detecting thyroid peroxidase antibodies based on nano magnetic particle chemiluminescence and alkaline phosphatase labeling. Background technique [0002] Thyroid peroxidase (TPO) is a membrane protein that binds glycosylated heme and is located in the apical membrane of thyroid follicular cells. TPO is a major component of thyroid microsomal antigen large protein, which acts as a catalyst in the iodination reaction of tyrosyl groups in thyroglobulin, and promotes the synthesis of thyroid hormones, T3 and T4. The detection of thyroid peroxidase autoantibodies is an effective means to identify whether a patient has autoimmune thyroid disease. More than 90% of patients with autoimmune thyroiditis have very high levels of anti-TPO antibodies. Anti-TPO antibodies can activate complement and are considered to be the main cause of thyroid dysfunct...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/553G01N33/531G01N21/76
CPCG01N33/577G01N21/76G01N33/531G01N33/553
Inventor 徐顺澍李永红周超高金艳吴荣桂李庆春
Owner SUZHOU HAOOUBO BIOPHARML
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