Human thyrotropin receptor antibody chemiluminescence detection kit as well as preparation method and application method thereof

A technology for chemiluminescent detection and thyrotropin detection, which is applied in the field of immunoassay medical detection, can solve the problems of inability to distinguish stimulatory, inhibitory, and neutralizing antibodies of human thyrotropin receptors, poor detection sensitivity, and poor accuracy. Securing Sensitivity, Enhancing Specificity, and Improving Sensitivity

Inactive Publication Date: 2018-12-14
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The present invention aims to solve the technical problem that the immunoassay method for detecting human thyrotropin receptor antibody in the prior art cannot distinguish stimulating, inhibitory and neutralizing

Method used

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  • Human thyrotropin receptor antibody chemiluminescence detection kit as well as preparation method and application method thereof
  • Human thyrotropin receptor antibody chemiluminescence detection kit as well as preparation method and application method thereof
  • Human thyrotropin receptor antibody chemiluminescence detection kit as well as preparation method and application method thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0042] The present invention also provides a preparation method of a human thyrotropin receptor antibody chemiluminescent detection kit, comprising the following steps:

[0043] 1) Preparation of calibrator with human thyrotropin stimulating antibody in human serum matrix;

[0044] 2) Preparation of quality control products with human thyrotropin-stimulating antibodies in human serum matrix;

[0045] 3) Preparation of R1 reagent

[0046] After diluting the streptavidin magnetic beads with the magnetic bead dilution buffer, prepare a streptavidin-coated magnetic bead solution with a magnetic bead concentration of 0.05%-0.1%, which is the R1 reagent;

[0047] 4) Preparation of R2 reagent

[0048] Use a 50KDa pore size ultrafiltration centrifuge tube to replace the thyrotropin receptor stimulating antibody buffer with 50mM-200mM PBS buffer, add acridinium ester, mix well, and place it at room temperature for 2-4 hours, then purify the marker with AKTA protein purifier Antibody...

Embodiment 1

[0054] Embodiment 1 prepares TRAb chemiluminescent detection kit of the present invention

[0055] 1. Preparation of calibrator

[0056] Dilute TRAb into calibrators with 100mM PBS buffer containing 20% ​​human serum, aliquot into 0mIU / mL, 0.3mIU / mL, 2.5mIU / mL, 5mIU / mL, 10mIU / mL, 20mIU / mL, 40mIU / mL .

[0057] 2. Preparation of Quality Control Products

[0058] The TRAb antibody was diluted with 100mM PBS buffer solution containing 20% ​​human serum to form a quality control product, and aliquoted into two-point quality control products with a concentration of 1mIU / mL and 25mIU / mL.

[0059] 3. Preparation of R1 reagent in magnetic bead solution coated with streptavidin

[0060] Dilute the streptavidin magnetic beads with magnetic bead dilution buffer to a concentration of 0.1%, and store at 4°C.

[0061] 4. Preparation of acridinium ester-labeled thyrotropin receptor stimulating antibody R2 reagent

[0062] Add 500 μg of thyrotropin receptor stimulating antibody into a 50K...

Embodiment 2

[0072] Embodiment 2 The methodological examination of TRAb chemiluminescence detection kit of the present invention

[0073] The test kit prepared in Example 1 is tested according to industry standards and testing procedures in the art, and the results are as follows:

[0074] 1. Determination of the linear range of the kit

[0075] One batch of the kit prepared in Example 1 was used to measure the linear range samples 0mIU / mL, 0.3mIU / mL, 2.5mIU / mL, 5mIU / mL, 10mIU / mL, 20mIU / mL, 40mIU / mL, and the linear The range is 0.3-40mIU / mL, the correlation coefficient R2=0.9999.

[0076] Table 1 Determination of the linear range of the kit

[0077] Determination of calibrator concentration (mIU / mL)

Relative Luminous Intensity (RLU)

0

142648

0.3

125453

2.5

65633

5

38415

10

18752

20

10247

40

5410

[0078] 2. Determination of Kit Sensitivity

[0079] With one batch of the kit prepared in Example 1, measure 2...

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Abstract

The invention relates to a human thyrotropin receptor antibody chemiluminescence detection kit as well as a preparation method and an application method thereof, and belongs to the technical field ofimmunoassay medical detection. The method can be used for solving the problems that an existing immunoassay method for detecting the human thyrotropin receptor antibody cannot distinguish stimulating,inhibition and neutralizing antibodies of human thyrotropin receptors, and the detection sensitivity and accuracy are poor. The kit disclosed by the invention uses a streptavidin-biotin system at thecombinational ratio of streptavidin to biotin of 1:4, so that the signal is further amplified, and the reagent sensitivity is improved. Specificity detection of the stimulating antibodies of the thyrotropin receptor is carried out by changing the marked antibody and the antigen, so that detection specificity is enhanced and more accurate and more targeted in-vitro diagnosis reagents are providedfor clinical diagnosis. The kit adopts a one-step competition method reaction mode, and the content of the stimulating antibodies of the thyrotropin receptor in human serum is quantitatively detected,so that the sensitivity and specificity of detection are ensured.

Description

technical field [0001] The invention relates to the technical field of immunoanalysis medical detection, in particular to a chemiluminescence detection kit for human thyrotropin receptor antibody and its preparation method and use method, which are used for the quantification of thyrotropin receptor stimulating antibody in human body detection. Background technique [0002] Thyroid-stimulating hormone receptor autoantibody (TRAb) is a group of polyclonal antibodies with heterogeneity. There are three main types: (1) Thyroid-stimulating antibody (TSAb), which can mimic the action of thyroid-stimulating hormone (TSH) to stimulate thyroid cells function and growth of the thyroid, causing hyperthyroidism and goiter. (2) Thyroid blocking antibody (TBAb), which can block the excitatory effect of TSH or TSAb on the thyroid gland, can cause addition and subtraction. (3) Neutralizing autoantibodies, which neither activate TSHR nor block the activation effect of TSAb on TSHR after b...

Claims

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Application Information

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IPC IPC(8): G01N33/76G01N33/68G01N33/58G01N33/532
CPCG01N33/532G01N33/582G01N33/583G01N33/6854G01N33/76G01N2800/046
Inventor 孙天旭李婷孙成艳高威何浩会
Owner DIRUI MEDICAL TECH CO LTD
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