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Development of Real Time RT-PCR Assays for Detection of Type A Influenza Virus and for Subtyping of Avian H5 and H7 Hemagglutinin Subtypes

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Abstract

Rapid detection and subtyping of H5 and H7 subtypes influenza A viruses are important for disease control in poultry and potential transmission to humans. Currently, virus isolation and subsequent HA and NA subtyping constitute the standard for avian influenza viruses detection and subtype identification. These methods are highly accurate and sensitive but are also laborious and time-consuming. Reverse transcription PCR and real time reverse transcription PCR assays, suitable tests for rapid detection, have previously been used for the specific diagnosis of H5 and H7 viruses, however, at present, no primer and probe sets are available for the identification of all H5 and H7 strains. Herein, we have developed specific and sensitive real time reverse transcription PCR assays for the detection of type A influenza virus and for subtyping of avian H5 and H7 hemagglutinin subtypes and we have also compared these molecular assays with viral isolation in terms of sensitivity. Our results demonstrate that the real time reverse transcription PCR assays are more sensitive, specific, less expensive compared to viral isolation. In conclusion, molecular assays could represent an useful tool for rapid detection and screening of H5 and H7 isolates during influenza A virus outbreaks alternatively to viral isolation.

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Acknowledgments

This work was supported by Fondo Regionale CIPE #A164. We want to thank National Reference Laboratory for Newcastle Disease and Avian Influenza, I.Z.S. Venezie for the supply of material.

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Authors and Affiliations

  1. Department of Public Health and Microbiology, Virology Unit, University of Turin, Via Santena 9, 10126, Torino, Italy

    Francesca Sidoti, Cristina Costa, Sara Astegiano, Antonio Curtoni, Rossana Cavallo & Massimiliano Bergallo

  2. Department of Virology, Veterinary Medical Research Institute for Piemonte, Liguria and Valle d’Aosta, Turin, Italy

    Francesca Rizzo & Maria Lucia Mandola

Authors
  1. Francesca Sidoti
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  2. Francesca Rizzo
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  3. Cristina Costa
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  4. Sara Astegiano
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  5. Antonio Curtoni
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  6. Maria Lucia Mandola
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  7. Rossana Cavallo
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  8. Massimiliano Bergallo
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Correspondence to Cristina Costa.

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Sidoti, F., Rizzo, F., Costa, C. et al. Development of Real Time RT-PCR Assays for Detection of Type A Influenza Virus and for Subtyping of Avian H5 and H7 Hemagglutinin Subtypes. Mol Biotechnol 44, 41–50 (2010). https://doi.org/10.1007/s12033-009-9211-7

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  • DOI: https://doi.org/10.1007/s12033-009-9211-7

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