Preparation method for quantum-dot-marked immunochromatographic 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
CHINA BEIJING BEIDA JUBANG SCI & TECH CO LTD
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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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  • Preparation method for quantum-dot-marked immunochromatographic test paper
  • Preparation method for quantum-dot-marked immunochromatographic 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 C 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 C virus monoclonal antibody, nitrocellulose membrane, and absorbent paper are pasted on the plastic plate in sequence;

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

[0035] Wherein, the quantum dot-labeled hepatitis C virus monoclonal antibody is located at the other end of the glass cellulose membrane B, corresponding to the detection band T and the qualit...

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 C 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 hepatitis C virus monoclonal antibody 150-200uL;

[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 C virus polyclonal antibody and rabbit anti-mouse secondary antibody with 0.05-0.15M PBS buffer, spray 0.5g / L hepatitis C virus polyclonal antibody and 1.0g / L rabbit anti-mouse secondary ...

Embodiment 3

[0058] Embodiment 3: detect hepatitis C virus monoclonal antibody with described test paper, comprise the following steps: sample point sample is approached one end of hepatitis C virus monoclonal antibody on the assembled test paper, after reacting for 5min, analyze in ultraviolet Observation results in the instrument. 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 detecting hepatitis c virus by quantum-dot-marked immunochromatographic test paper through immunology. According to the quantum-dot-marked immunochromatographic test paper, a glass cellulose membrane A, a glass cellulose membrane B of a quantum-dot-marked hepatitis c virus monoclonal antibody, a nitrocellulose membrane and absorbent paper are sequentially adhered to a plastic plate from top to bottom, wherein one end of the nitrocellulose membrane is provided with a hepatitis c virus polyclonal antibody and a rabbit antimouse second antibody to form a detection band T and a detection band C; the quantum-dot-marked hepatitis c virus monoclonal antibody is located at one end of the glass cellulose membrane B and corresponds to the detection band T and the detection band C, and the quantum-dot-marked hepatitis c virus monoclonal antibody is located at the end of a sampling point. The sensitiveness of the method is about 1000 times higher than that of a colloidal gold method.

Description

technical field [0001] The invention relates to a medical immunological detection method, in particular to a method for immunologically detecting hepatitis C virus by using quantum dot-labeled immunochromatographic test paper. Background technique [0002] Hepatitis C virus (HCV) infection is a worldwide epidemic disease, with more than 200 million chronically infected people worldwide. The clinical manifestations of HCV infection are various, ranging from mild inflammation, extensive liver fibrosis, liver cirrhosis, with or without hepatocellular carcinoma (hepatocellular carcinoma, HCC). [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. Therefore, it is an urgent problem to seek a detection method with low price, easy operation, high sensitivity and specificity. Contents of the invention [0004] Aiming at the shortcomings of the above ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/532
CPCG01N33/5767
Inventor 文德敏申有长于晓永
Owner CHINA BEIJING BEIDA JUBANG SCI & TECH CO LTD
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