title: A simple method for molecular detection of Swine-origin and human-origin influenza a virus.
abstract: Here we present a real-time one-step reverse transcriptase-polymerase chain reaction SYBR Green assay derived from the method reported by van Elden and collaborators ( 2001 ) that ensures the rapid, sensitive, and cost-effective detection of both seasonal influenza A virus and emerging (H1N1) swine-origin influenza A virus (S-OIV). In addition to this screening test, which successfully detected both seasonal influenza A virus and S-OIV in human clinical samples, we showed that the probe initially designed by van Elden and collaborators could detect seasonal influenza A virus, but not S-OIV; a new probe was designed and tested that specifically detects S-OIV, but not seasonal influenza A. Both probe-based assays were validated by testing human clinical samples and specifically detected either seasonal influenza virus or S-OIV. Finally, in silico analysis of databases predicted that minor modifications of the van Elden primers would facilitate the use of this assay for the broad spectrum detection of all currently characterized variants of influenza A virus, including avian strains.
authors.affiliation: Unité des Virus Emergents, Faculté de Médecine de Marseille, Université de la Méditerranée & Institut de Recherche pour le Développement, Marseille, France.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Ninove
authors.forename: Céline
authors.initials: C
authors.lastname: Gazin
authors.forename: Ernest A
authors.initials: EA
authors.lastname: Gould
authors.forename: Antoine
authors.initials: A
authors.lastname: Nougairede
authors.forename: Antoine
authors.initials: A
authors.lastname: Flahault
authors.forename: Rémi N
authors.initials: RN
authors.lastname: Charrel
authors.forename: Christine
authors.initials: C
authors.lastname: Zandotti
authors.forename: Xavier
authors.initials: X
authors.lastname: de Lamballerie
mesh_terms: D001483:Base Sequence; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics*; D009980:Influenza A virus / Q000235:genetics*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000191:economics / Q000379:methods / Q000592:standards; D012680:Sensitivity and Specificity; D016415:Sequence Alignment