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Method for detecting mumps virus, quantum-dot labeled immunochromatography test paper and preparation method thereof

A technology of immunochromatographic test paper and mumps virus, which is applied in the field of medical immunological detection, can solve the problems of low accuracy and low sensitivity, and achieve the effect of narrow emission peak, good luminescence stability and symmetrical peak shape

Active Publication Date: 2014-01-22
北京华卫骥生物医药有限公司
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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 detecting mumps virus, quantum-dot labeled immunochromatography test paper and preparation method thereof
  • Method for detecting mumps virus, quantum-dot labeled immunochromatography test paper and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: a quantum dot-labeled immunochromatographic test paper, provided with a plastic plate, a nitrocellulose membrane, a glass cellulose membrane A, a quantum dot-labeled glass cellulose membrane B for mumps virus IgG monoclonal antibody, and absorbent paper, The glass cellulose film A is a commercially available glass cellulose film without spotting;

[0033] Wherein, glass cellulose membrane A, glass cellulose membrane B marked with mumps virus IgG monoclonal antibody by quantum dots, nitrocellulose membrane, and absorbent paper are pasted on the plastic plate in sequence;

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

[0035] Wherein, the quantum dot-labeled mumps virus IgG monoclonal antibody is located at one end of the glass cellulose membrane B, corresponding to the detection band T and the quality control band C, and th...

Embodiment 2

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

[0042] (1) Conjugation of quantum dots with mumps virus IgG monoclonal antibody:

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

[0044] Select a coupling reagent, and the coupling reagent is selected from hydroxythiosuccinimide, 1-(3-dimethylaminopropyl)-3ethylcarbodiamine hydrochloride;

[0045] Add 150-200uL of mumps virus IgG monoclonal antibody;

[0046] Shaker reaction for 1 to 4 hours;

[0047] Chromatography 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 mumps virus polyclonal antibody and rabbit anti-mouse secondary antibody with 0.05-0.15M PBS buffer, spray 0.5g / L mumps virus polyclonal antibody and 1.0g / L rabbit anti-mouse secondary antibody o...

Embodiment 3

[0058] Embodiment 3: detect mumps virus with described test paper, comprise the following steps: spot sample on the assembled test paper close to one end of mumps virus IgG monoclonal antibody, after reaction 5min, observe result in ultraviolet analyzer . 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 particularly relates to a method for using quantum-dot labeled immunochromatography test paper to detect mumps 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 mumps 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 mumps 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 mumps 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 mumps virus IgG monoclonal antibody is positioned at one end of a sampling point. The method has the advantage that the detection sensitivity is about 1000 times higher than that of the commonly-used method.

Description

technical field [0001] The present invention relates to a medical immunological detection method, in particular to a method for detecting mumps virus by immunological method using quantum dots to mark immunochromatographic test paper. Background technique [0002] Mumps virus is the causative agent of mumps and belongs to the family Paramyxoviridae. The mumps virus is spherical, with a helical symmetric nucleocapsid and an envelope. The envelope has hemagglutinin-neuraminidase spikes (HN) and fusion factor spikes (F). The genome is single-stranded RNA. The mumps virus can proliferate in the amniotic cavity of chicken embryos or in chicken embryo cells, and cell fusion can occur, but the cytopathic changes are not obvious. There is only one serotype of mumps virus. It has weak resistance and can be inactivated in 30 minutes at 56°C, and is sensitive to ultraviolet rays and lipid solvents. Humans are the only host of the mumps virus, and the virus is spread by droplets. Th...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/577G01N2333/12
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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