Kit for detecting concentration of stimulating thyrotrophin receptor antibody

A thyroid-stimulating hormone and detection kit technology is applied in the field of detection kits for stimulating thyroid-stimulating hormone receptor antibody concentration, which can solve the problems of large error in experimental results, long detection time, complicated operation and the like, and achieves a wide detection range, High sensitivity and improved detection efficiency

Inactive Publication Date: 2019-12-17
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has many disadvantages, such as long detection time, cumbersome operation, low efficiency, and it is easy to cause large errors in experimental results.

Method used

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  • Kit for detecting concentration of stimulating thyrotrophin receptor antibody
  • Kit for detecting concentration of stimulating thyrotrophin receptor antibody
  • Kit for detecting concentration of stimulating thyrotrophin receptor antibody

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0034] In the embodiment of the present invention, the preparation method of the kit of the present invention comprises:

[0035] The first step, the preparation of magnetic particle suspension coated with stimulating thyroid-stimulating hormone receptor antibody

[0036] Take carboxyl magnetic particles according to the amount used, and activate them under acidic conditions with EDC and NHS. After the activation is completed, apply a magnetic field and let it stand to separate the magnetic particles from the liquid. Discard the supernatant and buffer with 0.01M PBS with pH 7.6. Wash away the excess activator with liquid; add an appropriate amount of stimulating thyroid-stimulating hormone receptor antibody, and shake the reaction under weak alkaline conditions; after the reaction, add a magnetic field and let it stand to separate the magnetic particles from the liquid. 0.01M PBS buffer solution of 1% bovine serum albumin for blocking, after the blocking, add blocking solution...

Embodiment 1

[0050] Embodiment 1 reagent preparation

[0051] The reagent preparation method of this embodiment comprises the following steps:

[0052] 1. Preparation of magnetic particle suspension coated with stimulating thyroid-stimulating hormone receptor antibody:

[0053] Take an appropriate amount of carboxyl magnetic particles according to the use, and activate them under acidic conditions (pH4.5-pH5.5) with excess EDC and NHS, and the activation buffer is 0.05M-0.1M MES (2-(N-? pheno)ethanesulfonic acid) buffer solution, the activation time is 30min; after the activation is completed, apply a magnetic field, and let it stand for 5min to separate the magnetic particles from the liquid; discard the supernatant, and wash twice with 0.01M PBS buffer solution of pH 7.6 , to wash away the excess activator; add an appropriate amount of stimulating thyroid stimulating hormone receptor antibody, so that the concentration is 22μg / mg magnetic particles, in 0.05M-0.1M MES buffer (pH4.5-pH5.5...

Embodiment 2

[0060] Embodiment 2 detection method

[0061] 1. Take out a certain amount of reaction containers (reaction wells) and number them, and add 50 μL of clinical serum samples or calibrators to each reaction well according to the experimental requirements;

[0062] 2. Shake the magnetic particle suspension, add 20 μL to each well;

[0063] 3. Add 50 μL of enzyme markers to each well;

[0064] 4. Mix the solution in the reaction well evenly and incubate at 37°C for 30 minutes;

[0065] 5. Using magnetic separation and washing equipment, wash the magnetic particles in the reaction well with washing solution for 5 times;

[0066] 6. Fully shake the washed reaction well to disperse the magnetic particles;

[0067] 7. Add 50 μL each of Luminescent Substrate A and Luminescent Substrate B to each well, shake and mix well, and keep away from light as a reaction for 5 minutes;

[0068] 8. Use a chemiluminescence detection instrument to detect the luminous intensity;

[0069] 9. Adopt ...

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Abstract

The invention relates to the technical field of in-vitro diagnosis, in particular to a kit for detecting the concentration of a stimulating thyrotrophin receptor antibody. The kit comprises magnetic particles coated with a stimulating thyrotrophin receptor antibody, a stimulating thyrotrophin receptor containing a marker label, and a stimulating thyrotrophin receptor antibody calibration product.According to the invention, the stimulating thyrotrophin receptor is used for constructing a stimulating thyrotrophin receptor antibody kit for the first time and the stimulating thyrotrophin receptoris only bound with the stimulating thyrotrophin receptor antibody, thereby eliminating the influence on the measuring system by the inhibiting thyrotrophin receptor antibody and the neutral thyrotrophin receptor antibody. The construction of the kit detection method is based on the high-sensitivity chemiluminiscence determination technology and the high-specificity immune reaction is combined; and thus the method has advantages of high sensitivity, high specificity and wide detection range.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a detection kit for the concentration of stimulating thyroid-stimulating hormone receptor antibody. Background technique [0002] TRAb is a thyroid autoantibody, which is a heterogeneous specific immunoglobulin produced by B lymphocytes with the help of T lymphocytes and some cytokines. Thyroid-stimulating hormone receptor (TSHR) is its main antigen. Thyroid-stimulating hormone receptor (TSHR) is a protein-coupled macromolecular glycoprotein. Human thyrotropin receptor is one of the important molecular components of the thyroid gland, mainly present on the membrane of thyroid follicle cells (TEC). Under physiological conditions, thyroid-stimulating hormone (TSH) binds to TSHR on the TEC membrane, activates adenylyl cyclase and phospholipase C, and passes through cyclic adenosine monophosphate and 1,4,5-triphosphate inositol, ethylene diphosphate, respectively. Phthal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/543G01N33/68
CPCG01N33/531G01N33/54326G01N33/68
Inventor 赵艳芳靳增明付光宇吴学炜
Owner AUTOBIO DIAGNOSTICS CO LTD
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