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Kit for determining thyrotropin and preparation method of kit

A thyrotropin and kit technology, which is applied in the field of immunoassay, can solve the problems of expensive time-resolved fluorescence immunoassay, low sensitivity of enzyme-linked immunoassay, and too sensitive metal chelate, so as to reduce non-specific adsorption, Improved sensitivity and accuracy, stable luminous intensity

Inactive Publication Date: 2019-12-27
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods all have some disadvantages: (1) the radioactive elements of radioimmunoassay pollute the environment and have a short half-life, which brings inconvenience to operation and limits the application of radioimmunoassay; (2) electrochemiluminescence is dissolved in the reaction system Oxygen interference and is too sensitive to metal ions, EDTA or other anticoagulants, preservative metal chelates; (3) enzyme-linked immunoassay has defects such as low sensitivity and narrow detection range, which cannot meet clinical requirements; (4) Time-resolved fluorescent immunoassay requires expensive and complex instrumentation

Method used

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  • Kit for determining thyrotropin and preparation method of kit
  • Kit for determining thyrotropin and preparation method of kit
  • Kit for determining thyrotropin and preparation method of kit

Examples

Experimental program
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Effect test

Embodiment 1

[0047] The preparation of embodiment 1 thyrotropin assay kit

[0048] (1) Reagent R1

[0049] Take an appropriate amount of streptavidin magnetic particle solution (purchased from Beijing Boermai Biotechnology Co., Ltd., with a particle size of 1-3 μm), add 20 mM PB buffer and dilute to 5 mg / mL, and mix well for 2 min. Place on a magnetic separator until the supernatant is clear without turbidity, then discard the supernatant. After repeating the washing operation 3 times, use 20mM PB buffer solution to prepare the R1 reagent with a magnetic bead concentration of 0.1%, and store it at 2-8°C.

[0050] (2) Reagent R2

[0051] Take an appropriate amount of thyrotropin antibody, replace the thyrotropin antibody with 20mM PB buffer, centrifuge for 10s in a handheld centrifuge, dilute the antibody to 2.0mg / mL, and mix the balanced acridinium ester stock solution with a vortex mixer at 1500rpm 20s, centrifuge for 10s in a handheld centrifuge, add the acridinium ester stock solutio...

Embodiment 2

[0054] Embodiment 2 Preparation of thyrotropin assay kit

[0055] (1) Reagent R1

[0056] Take an appropriate amount of streptavidin magnetic particle solution (purchased from Beijing Boermai Biotechnology Co., Ltd., with a particle size of 1-3 μm), add 20 mM PB buffer and dilute to 5 mg / mL, and mix well for 1 min. Place on a magnetic separator until the supernatant is clear without turbidity, then discard the supernatant. After repeating the washing operation 3 times, use 20mM PB buffer solution to prepare the R1 reagent with a magnetic bead concentration of 0.07%, and store it at 2-8°C.

[0057] (2) Reagent R2

[0058] Take an appropriate amount of thyrotropin antibody, replace thyrotropin antibody with 20mM PB buffer, centrifuge for 10s in a handheld centrifuge, dilute the antibody to 1.0mg / mL, and mix the balanced luminol stock solution with a vortex mixer at 1500rpm 20s, centrifuge for 10s in a handheld centrifuge, add the luminol stock solution to the antibody process...

Embodiment 3

[0061] Embodiment 3 Preparation of thyrotropin assay kit

[0062] (1) Reagent R1

[0063] Take an appropriate amount of streptavidin magnetic particle solution (purchased from Beijing Boermai Biotechnology Co., Ltd., with a particle size of 1-3 μm), add 20 mM PB buffer and dilute to 5 mg / mL, and mix well for 2 min. Place on a magnetic separator until the supernatant is clear without turbidity, then discard the supernatant. After repeating the washing operation for 3 times, use 20mM PB buffer to prepare the R1 reagent with a magnetic bead concentration of 0.15%, and store it at 2-8°C.

[0064] (2) Reagent R2

[0065] Take an appropriate amount of thyrotropin antibody, replace the thyrotropin antibody with 20mM PB buffer, centrifuge for 10s in a handheld centrifuge, dilute the antibody to 2.0mg / mL, and mix the balanced isoluminol stock solution with a vortex mixer at 1500rpm Mix for 20s, centrifuge for 10s in a handheld centrifuge, add the isoluminol stock solution to the ant...

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Abstract

The invention discloses a kit for determining thyrotropin and a preparation method of the kit. The kit comprises the following reagents of: a reagent R1, including a streptavidin magnetic particle solution, a reagent R2 including a chemiluminescence marker-labelled thyrotropin antibody, and a reagent R3 including a biotin-labeled thyrotropin antibody, wherein the mass percent of streptavidin magnetic particles in the reagent R1 is 0.07-0.2% and the particle sizes are 1-3 microns; the molar ratio of the thyrotropin antibody to a chemiluminescence marker in the reagent R2 is 1:1.5 to 1:10; and the molar ratio of the thyrotropin antibody to biotin in the reagent R3 is 1:5 to 1:50. The kit has the advantages of being high in sensitivity, high in specificity, free of a catalyst, stable in labelled conjugate, free of radioisotope damage and contamination, short in luminous time, stable in luminous intensity, high in reaction speed and high in specificity, and does not affect binding of a to-be-determined object after being connected with the antibodies.

Description

technical field [0001] The invention relates to the technical field of immune analysis, in particular to a thyrotropin assay kit and a preparation method thereof. Background technique [0002] Thyroid-stimulating hormone (TSH, thyrotropin) is a protein with a molecular weight of 30kD, which consists of two subunits. It is produced in specific basophils in the anterior pituitary gland, and its secretion is biologically rhythmic. Its concentration is affected by thyrotropin-releasing hormone secreted by the pituitary gland and thyroxine T4 and triiodothyronine T3 in the blood. It can regulate the function of thyroid cells, affect thyroid hormone synthesis, glucose oxidation and protein synthesis. When thyroid function changes, TSH levels fluctuate significantly. Therefore, TSH is a very sensitive and specific parameter for testing thyroid function, especially suitable for early detection or exclusion of dysfunction of the hypothalamus-pituitary-thyroid central regulatory loo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/78G01N33/53G01N33/531G01N33/543
CPCG01N33/5302G01N33/531G01N33/54326G01N33/78
Inventor 刘婷婷刘子菱韩美玉王香琪阚洪晶曲玉
Owner DIRUI MEDICAL TECH CO LTD
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