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Rapid Detection of Influenza A Virus Using a SPR Biosensor

Author: WangHaiJing
Tutor: HeHongXuan；ZhaoBaoHua
School: Hebei Normal
Course: Microbiology
Keywords: SPR biosensor Influenza A virus Rapid detection
CLC: Q939.4-3
Type: Master's thesis
Year: 2010
Downloads: 95
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Abstract


The H5N1 subtype of highly pathogenic avian influenza is a serious threat to human and animal health. The H5N1 avian influenza virus in Hong Kong in 1997 spread to people events highlight the public health significance of the prevention and control of avian influenza. Existing avian influenza diagnostic methods, including the method of virus isolation, RT-PCR, NASBA, colloidal gold immunochromatographic technology. Each diagnostic methods have their advantages, but is limited by several unfavorable factors, such as high cost, long processing cycles and low sensitivity, especially in the quantitative analysis of the instant, the accuracy of content and viruses of the avian influenza virus subtype determination needs to be improved. SPR biosensor of the present study, real-time monitoring of the sensor chip surface antigen-antibody binding / dissociation reaction is fast, simple, sensitive, specific, and low cost, as existing avian influenza diagnostic methods useful supplement for detection samples or chorioallantoic fluid, the avian influenza virus in cell cultures. This study prepared a detection of H5 subtype influenza virus antigen antibody chip and a detection of influenza A virus antigen antibody chip, single-channel single-parameter surface plasmon resonance biosensor H5 subtype of avian influenza virus type A The rapid detection methods for influenza viruses has been studied, both can be used directly for the detection of the sample. Rapid detection of H5 subtype influenza virus research, analysis and comparison of the effect of four different detection methods of detection, in order to obtain the most sensitive and rapid detection methods, these four methods were protein A fixed antibody assay, competitive inhibition assay antibody sandwich assay and direct detection method. Analysis of the test results is found that: the virus sample with 1% Triton X-100 lysis after 90 min at ambient conditions, by direct detection method is more rapid compared with several other methods. The direct detection method works as follows: anti-H5N1 subtype of influenza virus hemagglutinin (HA) monoclonal antibody immobilized on the sensor chip surface to capture viral antigen in the sample fragment, obtained by the change in refractive index of monitoring the course of the reaction surface of the chip associated antigen fragment binding quantity data, to analyze the data after determining the existence and content of the sample in the test object. H5 subtype influenza virus detection range established by direct detection method 5ng/mL-500ng/mL. Conducted a study on the specificity of the antibody chips, stability, and specific detection of the antibody chips on two H5N1 subtype avian influenza virus has no binding effect of contaminating proteins, and the same as the H9N2 subtype of influenza A virus-free binding reaction. Rapid detection of the H5 subtype of influenza A virus to establish the basis of this study were prepared by the detection of influenza A virus antibody chip, preliminary exploration and rapid detection of influenza A virus confirmed influenza A virus detection chip The highly pathogenic H5 subtype of influenza A virus of high hairstyle H9 subtype has a high specificity.

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CLC: > Biological Sciences > Microbiology > Microbial taxonomy (System microbiology ) > Virus ( viral )
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