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Chemiluminiscence detection kit for 17alpha-hydroxyprogesterone and preparation method and application thereof

A technology for chemiluminescence detection and α-hydroxyprogesterone, which is applied in chemiluminescence/bioluminescence, analysis by making materials undergo chemical reactions, and measurement devices, can solve problems such as poor sensitivity, decreased affinity, and poor stability, and achieve Effects with a wide linear range of influence

Inactive Publication Date: 2016-06-08
SHENZHEN NEW INDS BIOMEDICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are three major disadvantages in this technical solution: (1) when using the above-mentioned competition method, both the labeled antigen and the antigen to be tested are in liquid phase, and have the same chance of binding to the solid-phase antibody. When the concentration of 17α-hydroxyprogesterone antigen in the sample to be tested When it is low, the sensitivity is poor; (2) the 17α-hydroxyprogesterone antigen is a small molecule, and the solid phase of the antibody may easily cause some binding sites to be covered or changed, making its detection susceptible to interference and decreased affinity; ( 3) The enzyme-linked immunosorbent assay method takes a long time to detect, it takes at least 6 hours, or even overnight reaction to complete the test, the efficiency is low, and because the enzyme is easily affected by external interference factors, such as temperature, time and material concentration, resulting in its stability Poor performance, low specificity and poor sensitivity

Method used

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  • Chemiluminiscence detection kit for 17alpha-hydroxyprogesterone and preparation method and application thereof
  • Chemiluminiscence detection kit for 17alpha-hydroxyprogesterone and preparation method and application thereof
  • Chemiluminiscence detection kit for 17alpha-hydroxyprogesterone and preparation method and application thereof

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preparation example Construction

[0071] The following is the preparation method of each component of the kit:

[0072] Preparation 1: Preparation of derivatives formed by 17α-hydroxyprogesterone antigen and protein carrier

[0073] Taking the bovine IgG protein carrier as an example, to prepare 17α-hydroxyprogesterone antigen-bovine IgG derivatives, the specific steps are as follows:

[0074] Dissolve 1.45mg of 17-OH progesterone-3-carboxymethyl oxime, 0.52mg of NHS, and 0.85mg of EDC in 174μL DMSO, react at room temperature for 2h, in 3.6mL of bovine IgG solution (36mg of bovine IgG dissolved in 0.1M sodium bicarbonate), and stir at room temperature After reacting for 2 hours, it was purified by G25 gel column (eluted with 0.1M PBS buffer at pH 7.4) to obtain a derivative obtained by coupling 17-hydroxyprogesterone and bovine IgG.

[0075] It should be noted that when other protein carriers are used, the preparation method of their derivatives is compared with the above, the only difference lies in the type...

Embodiment 1

[0138] A chemiluminescent detection kit for 17α-hydroxyprogesterone, comprising the following components:

[0139] 1) Component A: Magnetic ball suspension coated with 17α-hydroxyprogesterone antigen derivative, the 17α-hydroxyprogesterone antigen derivative is formed by coupling 17α-hydroxyprogesterone antigen with bovine IgG protein carrier, wherein: magnetic The working concentration of the ball: 1mg / ml, the working concentration of the 17α-hydroxyprogesterone antigen is 50ng / ml, and the working concentration of the bovine IgG protein carrier is 10μg / ml;

[0140] 2) Component B: ABEI-labeled 17α-hydroxyprogesterone monoclonal antibody solution, wherein the working concentration of ABEI is 0.3 mg / l, and the working concentration of 17α-hydroxyprogesterone monoclonal antibody is 300 ng / ml;

[0141] 3) Calibrator solution: a low-concentration calibrator solution with a concentration of 0.433ng / ml and a high-concentration calibrator solution with a concentration of 11.567ng / ml;...

Embodiment 2

[0167] A chemiluminescent detection kit for 17α-hydroxyprogesterone, comprising the following components:

[0168]1) Component A: Magnetic ball suspension coated with 17α-hydroxyprogesterone antigen derivative, which is formed by coupling 17α-hydroxyprogesterone antigen with bovine serum albumin (BSA) carrier , wherein: the working concentration of magnetic balls: 1mg / ml, the working concentration of 17α-hydroxyprogesterone antigen is 50ng / ml, and the working concentration of BSA carrier is 10μg / ml;

[0169] 2) Component B: ABEI solution labeled with 17α-hydroxyprogesterone monoclonal antibody, wherein the working concentration of ABEI is 0.3 mg / ml, and the working concentration of 17α-hydroxyprogesterone monoclonal antibody is 300 ng / ml.

[0170] 3) Calibrator solution: a low-concentration calibrator solution with a concentration of 0.433 ng / ml and a high-concentration calibrator solution with a concentration of 11.567 ng / ml.

[0171] Each of the above components contains BS...

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Abstract

The invention provides a chemiluminiscence detection kit for 17alpha-hydroxyprogesterone. The chemiluminiscence detection kit comprises a component A and a component B, wherein the component A is a magnetic sphere suspension coated by a 17alpha-hydroxyprogesterone antigen derivative, and the 17alpha-hydroxyprogesterone antigen derivative is coupled by a 17alpha-hydroxyprogesterone antigen and a protein carrier; the component B is a chemiluminiscence marker solution for marking a 17alpha-hydroxyprogesterone antibody. The chemiluminiscence detection kit has the advantage that according to the principle of competitive method, the content of 17alpha-hydroxyprogesterone in a sample can be accurately and sensitively detected.

Description

technical field [0001] The invention relates to the technical field of biological substance detection, in particular to a chemiluminescent detection kit for 17α-hydroxyprogesterone and a preparation method thereof, and a method and device for detecting 17α-hydroxyprogesterone using the kit. Background technique [0002] 17α-Hydroxyprogesterone (17α-Hydroxyprogesterone, 17α-OHP) is an important small molecular intermediate in the biosynthesis of adrenocortical hormone. The normal adrenal gland uses cholesterol as a raw material to synthesize three main types of hormones: glucocorticoids, mineralocorticoids, and sex hormones (androgen, estrogen, and progesterone), all of which are derivatives of cholesterol. The process is extremely complicated, and each step must go through a series of Enzyme catalysis. These enzymes include 21-hydroxylase, 11β-hydroxylase, 3β-steroid dehydrogenase, 17α-hydroxylase, and the like. 17α-hydroxyprogesterone can be converted from progesterone un...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/532G01N21/76
CPCG01N33/54333G01N21/763G01N33/532G01N33/54346G01N33/54353
Inventor 饶微余慧玲李婷华彭国涛罗凯杜凯
Owner SHENZHEN NEW INDS BIOMEDICAL ENG
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