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Anti-Mullerian hormone quantitative measurement reagent and method based on nanometer up-converting phosphor method

A technology for quantitative detection of Mullerian hormones, applied in biological testing, measuring devices, material inspection products, etc., can solve problems such as high background value, high maintenance and testing costs, and high price

Active Publication Date: 2017-11-10
BEIJING HOTGEN BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In European and American countries, the blood AMH concentration level has become the main routine indicator for evaluating women’s ovarian function. The commonly used detection methods are enzyme-linked immunoassay and electrochemiluminescence method; ELISA and colloidal gold method; electrochemiluminescence method is mainly Roche's AMH electrochemiluminescence detection system, ELISA and colloidal gold method are mainly such as the patented "human anti-Müllerian hormone enzyme immunoassay reagent, CN 104865391B" and "Colloidal Gold Immunochromatography Detection Kit and Method for Human Anti-Müllerian Hormone, CN 105527447 A", respectively, but the electrochemiluminescence method is a non-open system, expensive, and requires special training equipment Users, maintenance and testing costs are high, not suitable for large-scale promotion and use; ELISA is time-consuming, low sensitivity, high background value, easy to cause false positive and false negative results; colloidal gold detection speed is fast, suitable for fast POCT Detection, but low sensitivity, high background value, easy to cause false positive and false negative results, quantitative detection relies on color or grayscale comparison, mostly semi-quantitative detection, the detection results cannot be accurately quantified

Method used

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  • Anti-Mullerian hormone quantitative measurement reagent and method based on nanometer up-converting phosphor method
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  • Anti-Mullerian hormone quantitative measurement reagent and method based on nanometer up-converting phosphor method

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Effect test

Embodiment 1

[0023] Embodiment 1: The process for the production of AMH up-transfer luminescence kit is as follows:

[0024] (1) Coating: Dilute the AMH monoclonal antibody (from Xiamen University) to 1.5 mg / ml, as a T-line coating solution, and dilute goat anti-mouse IgG (purchased from Beijing Suolaibao Biotechnology Co., Ltd.) to 2 mg / ml ml, as the C-line coating solution; the T-line coating solution and the C-line coating solution are coated on the nitrocellulose membrane through a film spraying machine; dry to obtain the monoclonal antibody-coated membrane strip, that is, the analysis membrane.

[0025] (2) UCP-labeled monoclonal antibody: The UCP-labeled monoclonal antibody in this kit is mainly obtained by the following steps, details are as follows:

[0026] a) Weigh 10 mg of UCP particles and place them in a conical flask;

[0027] b) Add 10ml of 0.20M PB solution with pH=7.2;

[0028] c) Add 0.5 mg anti-AMH antibody to the UCP particle suspension, then add anhydrous glutaraldeh...

Embodiment 2

[0042] Embodiment 2: Components of AMH up-transfer luminescence quantitative detection kit:

[0043] 1. AMH-UPT rapid detection test strip (40 servings)

[0044] 2. Sample diluent (1 bottle, 45ml)

[0045] 3. Sample tubes (40 pieces)

[0046] 4. Instruction manual (1 copy)

Embodiment 3

[0047] Embodiment 3: The principle of AMH-UPT immunochromatography test strip

[0048] The principle of the test strip: The AMH-UPT rapid detection test strip adopts the double antibody sandwich immunochromatography method to quantitatively detect AMH in the sample. The test area (T) on the analysis membrane is pre-coated with anti-AMH monoclonal antibody, the quality control area (C) is pre-coated with goat anti-mouse IgG, and there is another strain of anti-AMH monoclonal antibody labeled with UCP particles on the reagent pad. During the reaction, AMH in the sample reacts with two strains of antibodies to form an anti-AMH antibody-AMH antigen-UCP-labeled anti-AMH antibody complex in the T zone. UCP particles emit visible light under infrared light excitation. proportional to the concentration. Goat anti-mouse-UCP labeled anti-AMH antibody complexes are formed in the C region regardless of the presence or absence of AMH in the sample.

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Abstract

The invention relates to an anti-Mullerian hormone quantitative measurement reagent and method based on a rare earth nanometer up-converting phosphor method. UCP particles in which the up-converting phosphor method is adopted are used as a biomarker, and an anti-AMH antibody is specifically marked. Through the adoption of the anti-Mullerian hormone quantitative measurement reagent and method based on the up-converting phosphor method, the content of AMH in a measured sample can be quickly, simply, quantitatively and accurately measured, and a detection basis is provided for evaluation of the female ovarian reserve function.

Description

technical field [0001] The invention relates to a preparation method of a quantitative detection reagent and kit for anti-Müllerian hormone (AMH) based on a rare earth nanometer up-transition luminescence method, which belongs to the technical field of biotechnology and medical equipment, and is mainly used for the detection of female ovaries. The reserve function is evaluated. Background technique [0002] Anti-Mullerian Hormone (AMH), first discovered in 1947, is a member of the transforming growth factor β superfamily, a dimeric glycoprotein composed of two identical 70KD subunits linked by disulfide bonds , the relative molecular weight is 140KD; the human AMH coding gene is located on the short arm of chromosome 19, with a size of 2.4-2.8kb and 5 exons. [0003] AMH is a good indicator for evaluating ovarian reserve function, and it is not affected by menstrual cycle, and can be detected at any time. The AMH content in blood can provide a new reference index for the di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74G01N33/577
CPCG01N33/577G01N33/74G01N2800/36
Inventor 杨晓莉乔雍江兵泽余韶华李艳召李靖高琦汪吉杰张宏蕊林长青
Owner BEIJING HOTGEN BIOTECH CO LTD
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