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International Journal of Poultry Science

Year: 2009 | Volume: 8 | Issue: 3 | Page No.: 260-263
DOI: 10.3923/ijps.2009.260.263
Rapid Detection of Highly Pathogenic Avian Influenza H5N1 Virus by TaqMan Reverse Transcriptase-Polymerase Chain Reaction
B. P. Shankar, R. N.S. Gowda, B. Pattnaik, B.H. Manjunath Prabhu, S. S. Patil and H. K. Pradhan

Abstract: Highly pathogenic Avian Influenza (AI) H5N1 viruses have been spreading from Asia since late 2003. Early detection and classification are paramount for control of the disease because these viruses are lethal to birds and have caused fatalities in humans. Here we describe a TaqMan Reverse Transcriptase-Polymerase Chain Reaction Assay for rapid detection of Avian Influenza virus and for H5 subtyping by targeting HA gene of AI viruses. The assay was highly sensitive than RT-PCR and virus isolation in chick embryos. In the present study all samples (field samples) which are positive for HI and RT-PCR were tested by using TaqMan Reverse Transcriptase-Polymerase Chain Reaction Assay for reconfirmation. AI viruses (H5N1) were detected from nine samples which are received from Maharashtra during Avian influenza outbreak in India in 2006. Real-Time PCR assays was also conducted for detection of viral genome in different organs of experimental infected chickens revealed presence of the virus in all organs with high virus concentration in brain, heart, intestine and cloaca. This test allows definitive confirmation of an AI virus as H5 within hours, which is crucial for rapid implementation of control measures in the event of an outbreak.

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How to cite this article
B. P. Shankar, R. N.S. Gowda, B. Pattnaik, B.H. Manjunath Prabhu, S. S. Patil and H. K. Pradhan, 2009. Rapid Detection of Highly Pathogenic Avian Influenza H5N1 Virus by TaqMan Reverse Transcriptase-Polymerase Chain Reaction. International Journal of Poultry Science, 8: 260-263.

Keywords: Avian Influenza, H5N1 and TaqMan-RTPCR

REFERENCES

  • Dorak, M.T., 2001. Real-time PCR. Methods, 25: 1-8.


  • Heine, H.G., L. Trinidad, P. Selleck and S. Lowther, 2007. Rapid detection of highly pathogenic avian influenza H5N1 virus by taqman reverse transcriptase-polymerase chain reaction. Avian Dis., 51: 370-372.
    PubMed    Direct Link    


  • Jackwood, M.W., D.A. Hilt and S.A. Callison, 2003. Detection of infectious bronchitis virus by real-time reverse transcriptase-polymerase chain reaction and identification of a quasispecies in the Beaudette strain. Avian Dis., 47: 718-724.
    Direct Link    


  • Lee, C.W., D.L. Suarez, T.M. Tumpey, H.W. Sung and Y.K. Kwon et al., 2005. Characterization of highly pathogenic H5N1 avian influenza A viruses isolated from South Korea. J. Virol., 79: 3692-3702.
    Direct Link    


  • Ng, E.K.O., P.K. Cheng, A.Y. Ng, T.L. Hoang and W.W. Lim, 2005. Influenza a H5N1 detection. Emerg. Infect. Dis., 11: 1303-1305.
    PubMed    Direct Link    


  • Spackman, E., D.A. Senne, T.J. Myers, L.L. Bulaga and L.P. Garber et al., 2002. Development of a real-time reverse transcriptase PCR assay for type A Influenza virus and the Avian H5 and H7 hemagglutinin subtypes. J. Clin. Microbiol., 40: 3256-3260.
    PubMed    Direct Link    


  • Spackman, E., D.A. Senne, L.L. Bulaga, T.J. Myers and M. Perdue et al., 2003. Development of real-time RT-PCR for the detection of avian influenza virus. Avian Dis., 47: 1079-1082.


  • Yuen, K.Y., P.K.S. Chan, M. Peiris, D.N.C. Tsang and T.L. Que et al., 1998. Clinical features and rapid viral diagnosis of human disease associated with avian influenza A H5N1 virus. Lancet, 351: 467-471.
    CrossRef    PubMed    Direct Link    


  • Payungporn, S., S. Chutinimitkul, A. Chaisingh, S. Damrongwantanapokin and C. Buranathai et al., 2006. Single step multiplex real-time RT-PCR for H5N1 influenza A virus detection. J. Virol. Methods, 131: 143-147.
    CrossRef    

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