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Inhibitor A detection kit and preparation method thereof

A detection kit and inhibin technology, applied in the field of immunodiagnosis, can solve problems such as narrow linear range, slow luminescent reaction speed, and low sensitivity

Inactive Publication Date: 2018-12-21
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide an inhibin A detection kit and a preparation method thereof in order to solve the problems of slow luminescent reaction speed, low sensitivity, narrow linear range and high cost of the existing inhibin A detection kit

Method used

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  • Inhibitor A detection kit and preparation method thereof
  • Inhibitor A detection kit and preparation method thereof
  • Inhibitor A detection kit and preparation method thereof

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preparation example Construction

[0041] The present invention also provides a preparation method of an inhibin A detection kit, the method comprising:

[0042] Step 1: Preparation of Streptavidinized Magnetic Beads Suspension

[0043] 1) After ferric chloride and ferrous chloride are dissolved and mixed in water, ammonia water is added and reacted for 3-5h at 50-70° C. to obtain magnetic microspheres; the moles of ferric chloride and ferrous chloride The ratio is preferably 2:1;

[0044] 2) Wash the magnetic microspheres obtained in step 1) and redisperse them in 50% ethanol, then add 0.5% PEG4000, 0.1% ammonia water and 0.1% ethyl orthosilicate in sequence, and stir at 60-65°C for 14h-18h to obtain Modified magnetic microspheres; the volume ratio of the PEG4000, ammonia and tetraethyl orthosilicate is preferably 1:5:1;

[0045] 3) Wash the modified magnetic microspheres obtained in step 2) alternately with ultrapure water and absolute ethanol, resuspend them in 50% ethanol, and then add 0.05% ammonia water...

Embodiment 1

[0055] (1) Preparation of streptavidin-coated magnetic bead suspension

[0056] Weigh ferric chloride and ferrous chloride according to the molar ratio of 2:1, dissolve and mix them in ultrapure water, add ammonia water to a final concentration of 0.5%, stir and react at 60°C for 4 hours, and obtain magnetic microspheres;

[0057] Wash the obtained magnetic microspheres with ultrapure water and redisperse them in 50% ethanol, then add 1mL of 0.5% PEG4000, 5mL of 0.1% ammonia water and 1mL of 0.1% ethyl orthosilicate, and stir at 60°C for 16h. obtain modified magnetic microspheres;

[0058] The above-modified magnetic microspheres were washed alternately with ultrapure water and absolute ethanol, resuspended in 50% ethanol, and then 5 mL of 0.05% ammonia water, 2 mL of 0.5% tetramethylammonium hydroxide TMA and 4 mL of 2.5 % of 3-aminopropyltrimethoxysilane, stirred and reacted at room temperature for 16 hours, to obtain magnetic microspheres with surface-modified amino groups...

Embodiment 2

[0068] (1) Preparation of streptavidin-coated magnetic bead suspension

[0069] Weigh ferric chloride and ferrous chloride according to the molar ratio of 2:1, dissolve and mix them in ultrapure water, add ammonia water to a final concentration of 0.5%, stir and react at 60°C for 3 hours, and obtain magnetic microspheres;

[0070] Wash the obtained magnetic microspheres with ultrapure water and redisperse them in 50% ethanol, then add 1mL of 0.5% PEG4000, 5mL of 0.1% ammonia water and 1mL of 0.1% ethyl orthosilicate, and stir at 60°C for 14h. obtain modified magnetic microspheres;

[0071] The above-modified magnetic microspheres were washed alternately with ultrapure water and absolute ethanol, resuspended in 50% ethanol, and then 5 mL of 0.05% ammonia water, 2 mL of 0.5% tetramethylammonium hydroxide TMA and 4 mL of 2.5 % of 3-aminopropyltrimethoxysilane, stirring and reacting at room temperature for 14 hours, to obtain magnetic microspheres with surface-modified amino grou...

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Abstract

The invention provides an inhibitor A detection kit and a preparation method thereof to solve the problems that an existing inhibitor A detection kit is low in luminescent reaction rate, low in sensitivity, narrow in the linear range, and is high in the cost, and belongs to the technical field of immunodiagnosis. The kit comprises streptavidin magnetic bead suspension, an acridine-labeled inhibitor A antibody and biotin labeled inhibitor A antibody. The invention further provides a preparation method of the inhibitor A detection kit. The detection principle of a reagent is double antibody sandwich method. Detection is conducted by the acridine-labeled antibody and biotin labeled antibody. The luminous rate is increased and the reaction level is expanded. The specificity and detection speedof the reagent are greatly improved.

Description

technical field [0001] The invention belongs to the technical field of immunodiagnosis, and in particular relates to an inhibin A detection kit and a preparation method thereof. Background technique [0002] Inhibin is a heterodimeric glycoprotein hormone secreted by granulosa cells of the female ovary and Sertoli cells of the male testis. Inhibin selectively inhibits the secretion of follicle-stimulating hormone (FSH) from the pituitary gland, while also performing its own paracrine activities in the gonad. The fully processed inhibin molecule has a molecular weight of approximately 32 kD and consists of two distinct chains (α and β) covalently linked by a disulfide bond. The precursor is a high molecular weight form of the alpha subunit also found in flowing follicular fluid and serum. In addition, a free α-subunit form occurs, which is not linked to the β-subunit and which lacks the biological activity of inhibin. Inhibin A (INH-A / Inhibin A) includes an α subunit and a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74G01N33/531G01N33/543
Inventor 李冬梅高智玲高威孙成艳何浩会
Owner DIRUI MEDICAL TECH CO LTD
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