Preparation method for 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
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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  • Preparation method for quantum-dot labeled immunochromatography test paper
  • Preparation method for 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 mumps virus IgG 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, the plastic plate is pasted with glass cellulose membrane A, quantum dot-labeled mumps virus IgG monoclonal antibody glass cellulose membrane B, nitrocellulose membrane, and absorbent paper in sequence;

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

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

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 mumps 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 mumps 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 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 on nitro...

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 invention relates to a medical immunological detection method, in particular to a method for immunologically detecting mumps virus by using quantum dot-labeled immunochromatographic test paper. Background technique [0002] Adenovirus (mumps virus) is the causative agent of mumps, belonging to Paramyxoviridae. The mumps virus is spherical, the nucleocapsid is helically symmetrical, and has an envelope. There are hemagglutinin-neuraminidase spikes (HN) and fusion factor spikes (F) on the envelope. The genome is single negative-strand RNA. 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. The resistance is weak, it can be inactivated at 56°C for 30 minutes, and it is sensitive to ultraviolet rays and lipid solvents. Human beings are the only host of mumps virus, and the virus is transmit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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