Preparation method for quantum dot labelled immunochromatographic test strip

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
北京华卫骥生物医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 labelled immunochromatographic test strip
  • Preparation method for quantum dot labelled immunochromatographic test strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0033] Wherein, the plastic plate is pasted with glass cellulose membrane A, quantum dot-labeled glass cellulose membrane B of Japanese encephalitis virus IgG monoclonal antibody, nitrocellulose membrane, and absorbent paper in sequence;

[0034] Wherein, there are Japanese encephalitis 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 JEV IgG monoclonal antibody labeled with quantum dots is located at one end of the glass cellulose membrane B, correspond...

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 Japanese encephalitis virus IgG 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 150-200uL of Japanese encephalitis virus IgG 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 JEV polyclonal antibody and rabbit anti-mouse secondary antibody with 0.05-0.15M PBS buffer, spray 0.5g / L JEV polyclonal antibody and 1.0g / L rabbit anti-mouse secondary a...

Embodiment 3

[0058] Embodiment 3: detect the IgG monoclonal antibody of Japanese encephalitis virus with described test paper, comprise the following steps: spot sample on the assembled test paper near one end of IgG monoclonal antibody of Japanese encephalitis virus, react after 5min, analyze in ultraviolet Observation results in the instrument. PBS buffer solution and normal human blood were used as blank controls.

[0059] Result judgment: under 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 in particular relates to a method for detecting a Japanese encephalitis virus (JEV) by an immunological method by using a quantum dot labelled immunochromatographic test strip. The quantum dot labelled immunochromatographic test strip is characterized in that a glass cellulose membrane A, a glass cellulose membrane B of a quantum dot labelled JEV IgG (immunoglobulin G) monoclonal antibody, a nitrocellulose membrane and absorbent paper are stuck to a plastic board from bottom to top in sequence, wherein one end of the nitrocellulose membrane has a JEV polyclonal antibody and a rabbit anti-mouse second antibody, thereby forming a detection zone T and a quality control zone C; the quantum dot labelled JEV IgG monoclonal antibody is arranged at the other end of the glass cellulose membrane B, corresponds to the detection zone T and the quality control zone C and is arranged at one end of a sampling point. The detection sensitivity of the method is about 1000 times higher than that of the detection method frequently used at present.

Description

technical field [0001] The invention relates to a medical immunological detection method, in particular to a method for detecting Japanese encephalitis virus with an immunological method by using quantum dot-labeled immunochromatographic test paper. Background technique [0002] JEV was first isolated from the brain tissue of Japanese patients in 1935. Therefore, it is also called Japanese encephalitis virus (JEV). ~30nm, with a core composed of capsid protein and nucleic acid inside, covered with a lipid-containing capsule, with capsule glycoprotein spikes on the surface, that is, viral hemagglutinin, and an inner membrane protein in the capsule, which participates in the virus The total length of the JEV genome is 11kb, and it encodes structural proteins C, M, E and non-structural proteins NS1-NS5 sequentially from the 5' to 3' end. When the human body is bitten by a mosquito carrying the virus, the virus enters the blood circulation. The onset or not depends on the viru...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/533
CPCG01N33/56983G01N33/577G01N33/588G01N2333/185
Inventor 文德敏申有长于晓永
Owner 北京华卫骥生物医药有限公司
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