Thyroid microsomal antibody detection reagent kit as well as preparation method and application thereof

A technology of thyroid microsomes and kits, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of rare detection kits, and achieve the effects of long detection time, long storage time, and small system error

Active Publication Date: 2015-05-13
SHENZHEN NEW INDS BIOMEDICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, currently there are few detection kits with excellent per

Method used

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  • Thyroid microsomal antibody detection reagent kit as well as preparation method and application thereof
  • Thyroid microsomal antibody detection reagent kit as well as preparation method and application thereof
  • Thyroid microsomal antibody detection reagent kit as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0049] Example 1

[0050] (1) Labeling of protein A

[0051] Preparation of dialysate (F solution): add Na in a 5000ml beaker 2 CO 3 14.31g, NaHCO 3 26.46g, diluted with purified water to 4500ml. The prepared F solution was placed on a magnetic stirrer for use.

[0052] Select a dialysis bag with a suitable retention volume (usually 14000), measure the appropriate size, tie one end tightly after wetting, and test the purified water for leakage 3 times (no leakage is required).

[0053] Take 100 μg of protein A and adjust it to 1 ml with 0.1 mol / L pH9.5 carbonate buffer (F solution). Put it into the dialysate, stir and dialyze at room temperature for 2 hours, add 300 μg of ABEI activated ester to the dialyzed solution, and react at 37° C. for 2 hours.

[0054] The ligation product of protein A and ABEI was purified by G-25 gel column.

[0055] D 2 Preparation of solution: add 200ml of 0.5M phosphate buffer (P001 solution), 20g BSA, 8g NaN 3 , 2g MgCl 2 ·6H 2 O, 600ml...

Example Embodiment

[0072] Example 2

[0073] (1) Protein A biotinylation, the specific steps are as follows:

[0074] Preparation of dialysate (F solution): add Na in a 5000ml beaker 2 CO 3 14.31g, NaHCO 3 26.46g, diluted with purified water to 4500ml. The prepared F solution was placed on a magnetic stirrer for use.

[0075] 100 μg of biotin and 1 mg of protein A were taken and adjusted to 1 ml with 0.1 mol / L pH9.5 carbonate buffer (F solution). Put it into the dialysate, stir and dialyze at room temperature for 2 hours, and react at 37°C for 2 hours.

[0076] Purified by G-25 gel column.

[0077] The D2 solution was prepared according to the method of Example 1, and the purified ligation product was D2 2 The solution was diluted to a concentration of 0.025 μg / ml.

[0078] (2) Marking of SA

[0079] 100 μg of SA was adjusted to 1 ml with 0.1 mol / L carbonate buffer (F solution) of pH 9.5. Put it into the dialysate, stir and dialyze at room temperature for 2 hours, add 300 μg of ABEI ac...

Example Embodiment

[0095] Example 3

[0096] (1) Labeling of protein A

[0097] Preparation of dialysate (F solution): add Na in a 5000ml beaker 2 CO 3 14.31g, NaHCO 3 26.46g, diluted with purified water to 4500ml. The prepared F solution was placed on a magnetic stirrer for use.

[0098] Select a dialysis bag with a suitable retention volume (usually 14000), measure the appropriate size, tie one end tightly after wetting, and test the purified water for leakage 3 times (no leakage is required).

[0099] Take 100 μg of protein A and adjust it to 1 ml with 0.1 mol / L pH9.5 carbonate buffer (F solution). Put it into the dialysate, stir and dialyze at room temperature for 2 hours, add 300 μg of ABEI activated ester to the dialyzed solution, and react at 37° C. for 2 hours.

[0100] The ligation product of protein A and ABEI was purified by G-25 gel column.

[0101] D 2 Preparation of solution: add 200ml of 0.5M phosphate buffer (P001 solution), 20g BSA, 8g NaN 3 , 2g MgCl 2 ·6H 2 O, 600ml...

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Abstract

The invention relates to a chemical luminescent immunodetection thyroid microsomal antibody detection reagent kit and a preparation method of the reagent kit. The reagent kit comprises a protein A antigen labeled with a tracer marker, or an anti-human IgG and anti-human IgM labeled with the tracer marker, a thyroid microsomal antigen coated with magnetic spheres, or a junction complex of a protein carrier and the thyroid microsomal antigen coated with the magnetic spheres. The invention also provides a method for detecting the concentration of the thyroid microsomal antibody. When the reagent kit provided by the invention is used for detecting the concentration of the thyroid microsomal antibody, the enzymatic reaction is not involved, and the bridging of enzyme is not needed, so that the reagent is long in preservation time and stable; the detection also can be carried out by virtue of a full-automatic chemical luminescent method, so that the operation time is shortened, the manmade operation error is reduced, and the sensitivity and the accuracy in detection can be improved by utilizing the specificity of the chemical tracer marker.

Description

technical field [0001] The invention relates to a detection kit, in particular to a chemiluminescence immunoassay kit for detecting thyroid microsome antibody. The invention also relates to a preparation method of the kit and a method of using the kit to detect the concentration of thyroid microsome antibody. Background technique [0002] Thyroid microsomal antibody (TMA) is an antibody against thyroid microsomes (TM), which is one of the autoantibodies caused by autoimmune thyroid disease. TAM and antithyroglobulin antibody (TGA) have been recognized as important signs in the process of thyroid autoimmunity, and are the most representative antibodies. TAM and TGA are indispensable indicators in the diagnosis of autoimmune thyroid disease, and are one of the specific means for histological diagnosis of autoimmune thyroid disease. [0003] For various thyroid diseases, the positive detection rate of anti-TM: 50% to 100% for Hashimoto’s thyroiditis; 88.9% for hypothyroidism;...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/532G01N33/564G01N33/68
Inventor 饶微徐红陈帆李武杨雅丽李婷华袁锦云
Owner SHENZHEN NEW INDS BIOMEDICAL ENG
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