tetano
Editor, Senior Moderator
J Virol Methods. 2010 Nov 15. [Epub ahead of print]
Validation of commercial real-time RT-PCR kits for detection of Influenza A viruses in porcine samples and differentiation of pandemic (H1N1) 2009 virus in pigs.
Pol F, Qu?guiner S, Gorin S, Deblanc C, Simon G.
Anses, Laboratoire de Ploufragan-Plouzan?, Unit? Virologie Immunologie Porcines, Laboratoire National de R?f?rence pour les Virus Influenza Porcins, 22440 Ploufragan, France.
Abstract
Swine influenza, apart from its importance in animal health, may also be of public health significance. Although the first human infections with the multi-reassortant H1N1 virus (pH1N1/09) responsible for the 2009 pandemic were not related to pig exposure, this virus was shown to be related genetically to swine influenza viruses (SIV) and easily transmissible to pigs. In addition to direct animal health concerns, transmission and possible adaptation of the pH1N1/09 virus in pigs may have serious consequences on the risk of human infection by increasing the reservoir of this virus and the risk of possible emergence of new reassortant viruses with increased virulence for pigs and/or humans. Sensitive tools to monitor and detect rapidly such an infection are therefore mandatory. In this study, five commercial real-time RT-PCR assays developed by manufacturers LSI and Adiag?ne were assessed and validated, (i) for rapid detection of influenza A viruses, including pH1N1/09, in pig and (ii) for the differentiation of pH1N1/09 in that species. Two kits target the Influenza A virus M gene, two others amplify the pH1N1/09 virus H1 gene and one kit targets the pH1N1/09 virus N1 gene. All five kits are ready-to-use, one-step duplex RT-PCR and contain an internal positive control (IPC), appropriate for porcine biological samples, for assessing RNA extraction efficiency and the presence of PCR inhibitors. They have been used successfully by veterinary laboratories and shown to be powerful tools for the diagnosis and epidemiological surveillance of influenza virus infections in pigs.
Copyright ? 2010. Published by Elsevier B.V.
PMID: 21087637 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/21087637
Validation of commercial real-time RT-PCR kits for detection of Influenza A viruses in porcine samples and differentiation of pandemic (H1N1) 2009 virus in pigs.
Pol F, Qu?guiner S, Gorin S, Deblanc C, Simon G.
Anses, Laboratoire de Ploufragan-Plouzan?, Unit? Virologie Immunologie Porcines, Laboratoire National de R?f?rence pour les Virus Influenza Porcins, 22440 Ploufragan, France.
Abstract
Swine influenza, apart from its importance in animal health, may also be of public health significance. Although the first human infections with the multi-reassortant H1N1 virus (pH1N1/09) responsible for the 2009 pandemic were not related to pig exposure, this virus was shown to be related genetically to swine influenza viruses (SIV) and easily transmissible to pigs. In addition to direct animal health concerns, transmission and possible adaptation of the pH1N1/09 virus in pigs may have serious consequences on the risk of human infection by increasing the reservoir of this virus and the risk of possible emergence of new reassortant viruses with increased virulence for pigs and/or humans. Sensitive tools to monitor and detect rapidly such an infection are therefore mandatory. In this study, five commercial real-time RT-PCR assays developed by manufacturers LSI and Adiag?ne were assessed and validated, (i) for rapid detection of influenza A viruses, including pH1N1/09, in pig and (ii) for the differentiation of pH1N1/09 in that species. Two kits target the Influenza A virus M gene, two others amplify the pH1N1/09 virus H1 gene and one kit targets the pH1N1/09 virus N1 gene. All five kits are ready-to-use, one-step duplex RT-PCR and contain an internal positive control (IPC), appropriate for porcine biological samples, for assessing RNA extraction efficiency and the presence of PCR inhibitors. They have been used successfully by veterinary laboratories and shown to be powerful tools for the diagnosis and epidemiological surveillance of influenza virus infections in pigs.
Copyright ? 2010. Published by Elsevier B.V.
PMID: 21087637 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/21087637