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Comparison of Three Methods for the Detection of Avian Influenza Virus |
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Abstract: Three methods including the classical virus isolation and identification, reverse transcriptase polymerase chain reaction (RT-PCR) and indirect fluorescence assay (IFA) were used to detect avian influenza virus (AIV) in the sample and were compared. A virus was isolated by egg allantoic cavity inoculation and identified as H5 subtype of AIV with a monoclonal antibody (McAb) against H5 subtype via the hemoagglutination (HA) and hemoagglutination inhibit (HI) test. RT-PCR was performed to detect the sample directly and a 229bp-band of AIV specific and a 380bp-band of H5 subtype specific were both amplified with the designed primers. The tissue suspension was infected onto the monolayer of Madin-Darby canine kidney (MDCK) cells and a H5-McAb was applied to run the IFA on the monolayer cells at 24 hours post-infection. The fluorescence in both nucleolus and cytoplasm were observed. The results of the study demonstrated that these three methods could be applied to detect the AIV in the samp...更多le, while RT-PCR and IFA could identify the subtype of AIV at the same time, and have the advantages over the classical method in the aspects of sensitivity, rapidity and simplicity.
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Received: 24 April 2009
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