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Method for preparing quantum-dot labeled immunochromatography test paper

An immunochromatographic test strip and quantum dot technology, applied in the field of medical immunodetection, can solve the problems of low sensitivity and low accuracy, and achieve the effects of good luminescence stability, narrow emission peak and symmetrical peak shape.

Active Publication Date: 2015-07-22
BEIJING HUAWEI BRAVOBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the commonly used detection method is the colloidal gold method. Although this method is fast, simple and easy to operate, it has low accuracy and low sensitivity.

Method used

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  • Method for preparing quantum-dot labeled immunochromatography test paper
  • Method for preparing quantum-dot labeled immunochromatography test paper

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: a kind of quantum dot labeled immunochromatography test paper, is provided with plastic plate, nitrocellulose membrane, glass cellulose membrane A, quantum dot labeled hepatitis E virus monoclonal antibody glass cellulose membrane B, absorbent paper, Described glass cellulose film A is the glass cellulose film of buying on the market without spot;

[0033] Wherein, glass cellulose membrane A, quantum dot-labeled glass cellulose membrane B of hepatitis E virus monoclonal antibody, nitrocellulose membrane, and absorbent paper are pasted on the plastic plate in sequence;

[0034] Wherein, there are hepatitis E virus polyclonal antibody and rabbit anti-mouse secondary antibody at one end of the nitrocellulose membrane, so as to form detection zone T and quality control zone C;

[0035] Wherein, the hepatitis E virus monoclonal antibody labeled with quantum dots is located at one end of the glass cellulose membrane B, corresponding to the detection zone T and t...

Embodiment 2

[0041] Embodiment 2: the preparation method of test paper as mentioned above, as figure 1 shown, including the following steps:

[0042] (1) Coupling of quantum dots and hepatitis E virus monoclonal antibody:

[0043] Take 100-200uL of 0.01M PBS buffer and 5-20uL of quantum dots with carboxyl groups on the surface;

[0044] A coupling reagent is selected, and the coupling reagent is selected from hydroxysulfosuccinic acid imide, 1-(3-dimethylaminopropyl)-3 ethylcarbodiamine hydrochloride;

[0045] Add 150-200 uL of hepatitis E virus monoclonal antibody;

[0046] Shaker reaction for 1 to 4 hours;

[0047] Column filtration, centrifugal purification;

[0048] Block with 1% to 5% bovine serum albumin;

[0049] Store at 4°C;

[0050] (2) Preparation of test paper:

[0051] Dilute the hepatitis E virus polyclonal antibody and rabbit anti-mouse secondary antibody with 0.05-0.15M PBS buffer, spray 0.5g / L hepatitis E virus polyclonal antibody and 1.0g / L rabbit anti-mouse second...

Embodiment 3

[0058] Embodiment 3: detect hepatitis E virus with described test paper, comprise the steps: sample spot is approached one end of hepatitis E virus monoclonal antibody on the assembled test paper, after reaction 5min, observe in ultraviolet analyzer result. PBS buffer solution and normal human blood were used as blank controls.

[0059]Result judgment: On the premise that the C band shows a red fluorescent band, the intensity of the fluorescent band of the T band is visually compared with the blank. The weaker the fluorescence, the lower the concentration of the tested substance in the test solution.

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Abstract

The invention relates to a medical immunodetection method and particularly relates to a method for using quantum-dot labeled immunochromatography test paper to detect hepatitis E virus by an immunological method. The quantum-dot labeled immunochromatography test paper is characterized in that a plastic board is stuck with a glass cellulose membrane A, a glass cellulose membrane B of a quantum-dot labeled hepatitis E virus IgG monoclonal antibody, a nitrocellulose membrane and water absorbing paper from bottom to top in sequence, wherein one end of the nitrocellulose membrane is provided with hepatitis E virus polyclonal antibody and rabbit antimouse second antibody so as to form a detecting band T and a quality control band C; the quantum-dot labeled hepatitis E virus IgG monoclonal antibody is positioned at one end of the glass cellulose membrane and corresponds to the detecting band T and the quality control band C, and the quantum-dot labeled hepatitis E virus IgG monoclonal antibody is positioned at one end of a sampling point. The method has the advantages that high specificity of immunoreactions and the fluorescence characteristic of quantum dots are combined, so that the detection sensitivity is about 1000 times higher than that of the current commonly-used detection method.

Description

technical field [0001] The invention relates to a medical immunological detection method, in particular to a method for immunologically detecting hepatitis E virus by using quantum dot-labeled immunochromatographic test paper. Background technique [0002] Hepatitis E is a zoonotic disease of humans and various animals caused by the hepatitis E virus. It is mainly transmitted through the fecal route, and outbreaks are often caused by contaminated drinking water sources, and sporadic cases are distributed globally. Hepatitis E accounts for about 15% to 20% of acute hepatitis in my country. The mortality rate of adults suffering from hepatitis E is 0.5% to 3%, while that of pregnant women is as high as 15% to 20%. Therefore, the screening and diagnosis of hepatitis E is particularly important. [0003] At present, the commonly used detection method is the colloidal gold method. Although this method is fast, simple and easy to operate, it has low accuracy and low sensitivity...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577
CPCG01N33/576
Inventor 文德敏申有长于晓永
Owner BEIJING HUAWEI BRAVOBIO
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