Reagent kit and method for detecting thyroglobulin antibody

A thyroglobulin and kit technology, applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of environmental pollution, short validity period, high radioactivity, etc., and achieve the effect of overcoming high pollution and improving environmental performance.

Inactive Publication Date: 2016-02-17
SUZHOU HAOOUBO BIOPHARML
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Radioimmunoassay (RIA) has shortcomings such as high radioactivity, short validity period, health hazards to operators, and environmental pollution. Ordinary microplate enzyme-

Method used

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  • Reagent kit and method for detecting thyroglobulin antibody
  • Reagent kit and method for detecting thyroglobulin antibody

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Experimental program
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Effect test

Embodiment 1

[0025] Provided is a kit for detecting thyroglobulin antibody, comprising a first reagent, a second reagent, a magnetic separation reagent, a chemiluminescent substrate, a calibrator, a quality control and a cleaning solution, the first reagent being thyroglobulin ( Tg) labeled biotin reagent, the second reagent is thyroglobulin (Tg) enzyme conjugate reagent, and the magnetic separation reagent is streptavidin-labeled superparamagnetic nano-magnetic particle reagent.

[0026] The magnetic separation reagent includes: superparamagnetic nano-magnetic particles based on the surface of magnetic beads activated by toluenesulfonyl groups on the surface of hydrophobic magnetic beads, the diameter of the magnetic separation reagent is 1.05 μm, the isoelectric point is pH5.0, and at pH7, The charge is -10mV and the iron content is 26%. The superparamagnetic nano-magnetic particles are directly or indirectly labeled with streptavidin. The superparamagnetic nano-magnetic particles are n...

Embodiment 2

[0033] A method for detecting thyroglobulin antibody is provided, comprising the steps of:

[0034] (1) Add 10 μL of the sample to be tested, 50 μL of streptavidin-labeled superparamagnetic nanoparticle reagent and 100 μL of thyroglobulin (Tg)-labeled biotin reagent in the test tube, mix well and react at 37°C for 15 minutes.

[0035] (2) Add 500 μL of cleaning solution to wash away unbound antigens, antibodies and impurities, and repeat the washing 3 times.

[0036] (3) Add 100 μL of thyroglobulin (Tg) enzyme conjugate reagent, mix well and react at 37°C for 15 minutes.

[0037] (4) Add 500 μL of cleaning solution to wash away unbound antigens, antibodies and impurities, and repeat the washing 3 times.

[0038] (5) Add 150 μL of chemiluminescence substrate solution to each tube, react at 37°C for 5 minutes, and put it into a chemiluminescence analyzer / measurement instrument to measure the luminescence intensity (RLU).

[0039] After incubation in the above steps, unbound co...

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Abstract

The invention discloses a reagent kit and method for detecting a thyroglobulin antibody. The reagent kit contains a first reagent, a second reagent, a magnetic separation reagent, a chemical light-emitting substrate, a calibration article, a quality control article and cleaning liquid. The first reagent is a biotin reagent marked by thyroglobulin. The second reagent is a thyroglobulin enzyme combination reagent. The magnetic separation reagent is a superparamagnetic nanometer magnetic particle reagent marked by streptavidine, and the reagent kit is adopted for detecting the thyroglobulin antibody. Due to the mode, the defects that radioimmunoassay is high in pollution, the period of validity is short, common microwell plate enzyme immunoassay precision is poor, and sensitivity is poor are overcome, an alkaline phosphatase enzyme catalysis chemiluminescence system, a magnetic particle separation system and an unique antigen and antibody coupling technology are used, and the sensitivity, the precision, the stability, the validity period, the safety and the environment friendliness of the method are greatly improved.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a kit and method for detecting thyroglobulin antibody based on nano magnetic particle chemiluminescence and alkaline phosphatase labeling. Background technique [0002] Thyroglobulin (Tg) is a large heterologous glycoprotein (MW660,000) present in thyroid follicular cells. Thyroglobulin plays a very important role in the biosynthesis of thyroid hormones, T3 and T4. In thyroid follicle cells, thyroid peroxidase (TPO) acts as a catalyst for the iodination of tyrosyl groups in thyroglobulin. The iodinated thyroglobulin will be stored in the colloid of the follicle, acting as a depositor of T3 and T4. When the thyroid is stimulated, thyroglobulin is degraded and thyroid hormones, T3 and T4, are released into the blood. [0003] The detection of thyroglobulin autoantibodies is an effective means to identify whether patients suffer from autoimmune thyroid disease. Anti-T...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/535
CPCG01N33/53G01N33/535G01N2800/046
Inventor 周超李永红徐顺澍高金艳吴荣桂李庆春
Owner SUZHOU HAOOUBO BIOPHARML
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