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Method for detecting EB (Epstein-Barr) virus, quantum dot labelled immunochromatographic test strip and preparation method thereof

A technology of immunochromatography test paper and EB virus, which is applied in the field of medical immunological detection, 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: 2014-01-29
北京华卫天和生物科技有限公司
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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 EB (Epstein-Barr) virus, quantum dot labelled immunochromatographic test strip and preparation method thereof
  • Method for detecting EB (Epstein-Barr) virus, quantum dot labelled immunochromatographic test strip and preparation method thereof

Examples

Experimental program
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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 EB virus IgA monoclonal antibody, absorbent paper, so The above-mentioned glass cellulose film A is the glass cellulose film purchased on the market without sample;

[0033] Wherein, the glass cellulose membrane A, the glass cellulose membrane B of the EB virus IgA monoclonal antibody labeled with quantum dots, the nitrocellulose membrane, and the absorbent paper are pasted successively on the plastic plate;

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

[0035] Wherein, the Epstein-Barr virus IgA monoclonal antibody labeled with quantum dots is located at the other end of the glass cellulose membrane B, correspondin...

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 Epstein-Barr virus IgA 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 hydroxysulfosuccinimide, 1-(3-dimethylaminopropyl)-3 ethylcarbodiamine hydrochloride;

[0045] Add EB virus IgA 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 EB virus polyclonal antibody and rabbit anti-mouse secondary antibody with 0.05-0.15M PBS buffer, spray 0.5g / L EB virus polyclonal antibody and 1.0g / L rabbit anti-mouse secondary antibody on one end of the nit...

Embodiment 3

[0058]Embodiment 3: Detect EB virus with described test paper, comprise the following steps: spot sample on the assembled test paper close to one end of EB virus IgA monoclonal antibody, after reacting for 5min, observe the result in the 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 in particular relates to a method for detecting an EB (Epstein-Barr) virus 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 EB virus IgA (immunoglobulin A) 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 is provided with an EB virus 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 EB virus IgA 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 Epstein-Barr virus in an immunological way by using quantum dot-labeled immunochromatographic test paper. Background technique [0002] Epstein-Barr virus was the first time that Epstein and Barr successfully established Burkitt African children's lymphoma cells through in vitro suspension culture in 1964, and observed herpes virus particles in the cell smears of the established strain with electron microscopy, and believed that the virus was a variety of malignant One of the causes of tumors (such as nasopharyngeal carcinoma), which mainly infects the epithelial cells and B lymphocytes of the human oropharynx. Most of the patients with nasopharyngeal carcinoma in southern China have detected the presence of Epstein-Barr virus genome. The disease is widely distributed, mostly sporadic, and can also cause epidemics. Virus carriers and patients ar...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/533
CPCG01N33/533G01N33/56983G01N33/56994G01N33/577G01N2333/05
Inventor 文德敏申有长于晓永
Owner 北京华卫天和生物科技有限公司
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