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Chemiluminescence immune analytic reagent kit for detecting thyroid peroxidase autoantibody

A chemiluminescent immunoassay and peroxidase technology, applied in the field of immunoassay, can solve the problem of low sensitivity

Inactive Publication Date: 2009-03-04
北京科美东雅生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the method of direct chemiluminescence immunoassay is only used, the sensitivity of the method is relatively low, and the use of biotin-avidin system can avoid such shortcomings

Method used

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  • Chemiluminescence immune analytic reagent kit for detecting thyroid peroxidase autoantibody

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Preparation of TPOAb chemiluminescence immunoassay assay kit of the present invention

[0021] 1. Preparation of TPOAb calibrator

[0022] Pure TPOAb was diluted into calibrator with horse serum, the concentrations were 0, 25, 50, 150, 300, 600 IU / mL.

[0023] 2. Preparation of TPO antigen-coated microwell plates

[0024] a) coated

[0025] Mix 0.05M PBS buffer solution with pH7.2 and appropriate concentration of thyroid peroxidase antigen to make coating solution, and load it on a luminescent microwell plate;

[0026] b) washing the above-mentioned microwell plate with a washing solution; and

[0027] c) closed

[0028] Prepare hydrolyzed gelatin blocking solution, based on 1000mL of the blocking solution contains: 1.28mMNaH 2 PO 4 2H 2 O, 8.10 mM Na 2 HPO 4 12H 2 O, 1.0% hydrolyzed gelatin, 0.1% Proclin300, the pH is 7.2-7.4, and then the obtained blocking solution is loaded on the above-mentioned washed microporous plate for sealing for 2 hours, t...

Embodiment 2

[0062] Embodiment 2 The usage method of kit of the present invention

[0063] 1. Sample pretreatment

[0064] The fasting morning serum samples were taken from people, centrifuged at 3000rpm for 5min, and the upper serum was taken for analysis.

[0065] 2. Detection method

[0066] Follow the steps below to use this kit:

[0067] 1) Preparation of washing buffer: dilute the washing buffer provided in the kit with distilled water 25 times;

[0068] 2) Take the calibrator, coated microwell plate, biotin marker and enzyme marker out of the refrigerator at 4°C, place it for 20min, and equilibrate to room temperature;

[0069] 3) Add 0, 25, 50, 150, 300, 600 IU / mL of 0, 25, 50, 150, 300, 600 IU / mL to the reaction wells respectively, and add 1 blank well for each test, and then add Biotin labeling solution 100μL, react at 37℃ for 30min;

[0070] 4) Discard the solution in the well, wash 5 times with the diluted washing buffer, and pat dry on absorbent paper;

[0071] 5) Add 10...

Embodiment 3

[0077] Embodiment 3 The methodological test of the kit of the present invention

[0078] The test kit prepared in Example 1 is tested according to the conventional manufacturing and testing procedures in the art. The present invention has carried out specific experimental operations on the precision, accuracy, sensitivity, specificity and stability of the test kit. The experimental results are shown in Table 1. The results showed that the sensitivity, precision, accuracy, specificity and stability of the "Thyroid Peroxidase Autoantibody Chemiluminescent Immunoassay Assay Kit" were fully qualified.

[0079] Table 1 Test results of technical indicators of the kit

[0080] Test items Inspection standards test result accuracy The average recovery rate is 90.0~110.0% 94.2～108.6％ specificity Cross-reaction rate with its analogues ≤0.1% <0.07％

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Abstract

The invention provides a chemiluminescent immunoassay kit for thyroid peroxidase autoantibodies and a preparation method thereof. The invention has the advantages that the reaction pattern of the double antigen sandwich method is adopted, the chemiluminescent technology and the biotin-avidin immune amplifying technological principle are effectively utilized, horse radish peroxidase is adopted to be as the marker enzyme, the content of the thyroid peroxidase autoantibodies in human serum samples is quantitatively detected, and the detection sensitivity can be ensured. The kit of the invention has the advantages of high sensitivity, good repeatability, good stability and high detection efficiency, and can provide a timely and reliable experimental basis for the clinical diagnosis and the treatment of thyroid disorders.

Description

technical field [0001] The invention relates to the field of immune analysis, in particular to a thyroid peroxidase autoantibody assay kit and a preparation method thereof. The kit of the invention combines biotin-avidin immunomagnification technology and chemiluminescence immunoassay technology. Background technique [0002] The human thyroid is an important part of the endocrine system, and thyroid hormones participate in many important physiological reactions. They play an important regulatory role in human growth and development, cell metabolism and hormone balance in the body, as well as in maintaining metabolism, skeletal muscle and the development of various organ systems. Autoimmune thyroid disease (AITD) is an organ-specific autoimmune disease mediated by T cells. There are a variety of autoantibodies against thyroid antigens in the serum of patients. Thyroid peroxidase antibody (TPOAb) is One of the most important, prevalent in autoimmune thyroid disease. Studie...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/543G01N33/535G01N21/76
Inventor 辛天兵应希堂宋胜利胡国茂郑金来于尚永
Owner 北京科美东雅生物技术有限公司
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