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Detection micro particle of thyroid stimulating hormone and preparation and application thereof

A technology of thyroid stimulating hormone and microparticles, which can be used in biological tests, measuring devices, material inspection products, etc., and can solve the problems of expensive instruments and complex automation design.

Active Publication Date: 2014-07-30
BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the separation and cleaning involved in the reaction process, the automation design is complicated and the equipment is quite expensive
In addition, the stability of the luminescent matrix is ​​also a big problem

Method used

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  • Detection micro particle of thyroid stimulating hormone and preparation and application thereof
  • Detection micro particle of thyroid stimulating hormone and preparation and application thereof
  • Detection micro particle of thyroid stimulating hormone and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] Preparation of luminescent particles coated with TSHAb in embodiment 1

[0098] Preparation method of antibody and luminescent particles:

[0099] 1) Suspension treatment of luminescent particles: absorb a certain amount of carboxyl luminescent particles and centrifuge in a high-speed refrigerated centrifuge, discard the supernatant, add a certain amount of MES buffer, ultrasonically break until the particles are resuspended, add MES buffer to adjust the luminescent particles Concentration to 100mg / ml.

[0100] 2) Antibody treatment: TSH antibody was dialyzed against 0.05M MES buffer at pH 6.0 (hereinafter referred to as MES buffer). After the dialysis, the concentration was measured and adjusted to 8 mg / ml.

[0101] 3) MES buffer, 100mg / ml luminescent microparticle suspension (MES buffer) and 8mg / ml anti-TSH (MES buffer) and mixed at a volume ratio of 1:1:1, and mixed quickly , to obtain the reaction solution.

[0102] 4) Prepare 40mg / ml EDAC solution with MES buffe...

Embodiment 2

[0150] Example 2 Preparation of biotin-labeled antibody

[0151] Preparation:

[0152] 1) Antibody treatment: dialyze anti-TSH antibody in 0.1M NaHCO 3 solution, the antibody concentration was measured and adjusted to 1 mg / ml.

[0153] 2) Prepare 16.17mg / ml Biotin solution with DMSO.

[0154] 3) Labeling: Take the processed 1 mg / ml anti-AFP antibody labeled antibody and the prepared Biotin solution, mix them according to the volume ratio of 72:10000, and mix them uniformly quickly. Stand at 4°C for 12-16 hours.

[0155] 4) Dialysis: Dialyze the reacted biotin-labeled antibody against biotin-labeled dialysis buffer (pH 8.0).

[0156] 5) Aspirate the dialyzed biotinylated antibody and transfer it to a clean centrifuge tube, and take a sample to determine the antibody concentration. Adjust the concentration of qualified biotin-labeled antibody to 0.5mg / ml.

[0157] React antibodies with Biotin in different ratios and detect:

[0158] The detection method of each parameter ...

Embodiment 3

[0191] Example 3 Preparation of Photosensitive Microparticles Coated with Avidin

[0192] Photosensitive particles: use photosensitive particles with a particle size of 220±40nm (PentaTek, USA)

[0193] Preparation:

[0194] a. Treatment of photosensitive particle suspension: absorb a certain amount of photosensitive particles and centrifuge in a high-speed refrigerated centrifuge, discard the supernatant, add a certain amount of MES buffer, ultrasonically sonicate on the ultrasonic cell disruptor until the particles are resuspended, and then add MES buffer Adjust the photosensitive particle concentration to 100mg / ml.

[0195] b. Preparation of avidin solution: weigh a certain amount of Avidin, add MES buffer to dissolve to 8mg / ml.

[0196] c. Mixing: Mix the treated photosensitive microparticle suspension, 8 mg / ml Avidin and MES buffer at a volume ratio of 2:5:1, and mix quickly to obtain a reaction solution.

[0197] d. Reaction: Prepare 25mg / ml NaBH in MES buffer 3 CN s...

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Abstract

The invention relates to an aided diagnostic reagent of thyroid diseases and discloses a detection micro particle of thyroid stimulating hormone, which is a luminous micro particle coated by an antithyrotropic hormone antibody. The invention also discloses preparation and application of the thyroid stimulating hormone. In addition, the invention further discloses a vitro diagnostic kit of the thyroid stimulating hormone in a determined sample. Meanwhile, the invention discloses a using method of the kit. The kit can be combined with other serums and clinical information for aided diagnosis of the thyroid diseases and the monitoring of treatment effect of related patients.

Description

technical field [0001] The invention relates to a diagnostic reagent for thyroid-stimulating hormone, in particular to a microparticle for detecting thyroid-stimulating hormone based on the principle of light-induced chemiluminescence, its preparation and application. Background technique [0002] Thyroid disease is a group of more common endocrine diseases, including iodine deficiency goiter, goiter caused by other reasons, hyperthyroidism, thyroid tumor, thyroiditis, hypothyroidism and so on. The incidence of thyroid disease is relatively high. [0003] Hyperthyroidism, also known as hyperthyroidism, is a very common clinical endocrine disease. Refers to the enhancement of thyroid function caused by various reasons, excessive secretion of thyroid hormones or increased levels of thyronine (TSH) in the blood caused by the nervous system, circulatory system, digestive system, cardiovascular system and other multiple systems of the body. A range of hypermetabolic syndromes a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/78G01N33/543G01N33/52G01N21/76G01N21/64
Inventor 张伟王海蛟赵卫国
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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