title: Detection of novel influenza A(H1N1) virus by real-time RT-PCR.
abstract: Accurate and rapid diagnosis of novel influenza A(H1N1) infection is critical for minimising further spread through timely implementation of antiviral treatment and other public health based measures. In this study we developed two TaqMan-based reverse transcription PCR (RT-PCR) methods for the detection of novel influenza A(H1N1) virus targeting the haemagglutinin and neuraminidase genes. The assays were validated using 152 clinical respiratory samples, including 61 Influenza A positive samples, collected in Queenland, Australia during the years 2008 to 2009 and a further 12 seasonal H1N1 and H3N2 influenza A isolates collected from years 2000 to 2002. A wildtype swine H1N1 isolate was also tested. RNA from an influenza A(H1N1) virus isolate (Auckland, 2009) was used as a positive control. Overall, the results showed that the RT-PCR methods were suitable for sensitive and specific detection of novel influenza A(H1N1) RNA in human samples.
authors.affiliation: Queensland Paediatric Infectious Diseases Laboratory, Sir Albert Sakzewski Virus Research Centre, Queensland Children's Medical Research Institute, Children's Health Service District, Queensland, Australia. d.whiley@uq.edu.au
authors.forename: David M
authors.initials: DM
authors.lastname: Whiley
authors.forename: Seweryn
authors.initials: S
authors.lastname: Bialasiewicz
authors.forename: Cheryl
authors.initials: C
authors.lastname: Bletchly
authors.forename: Cassandra E
authors.initials: CE
authors.lastname: Faux
authors.forename: Bruce
authors.initials: B
authors.lastname: Harrower
authors.forename: Allan R
authors.initials: AR
authors.lastname: Gould
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Lambert
authors.forename: Graeme R
authors.initials: GR
authors.lastname: Nimmo
authors.forename: Michael D
authors.initials: MD
authors.lastname: Nissen
authors.forename: Theo P
authors.initials: TP
authors.lastname: Sloots
mesh_terms: D000818:Animals; D001315:Australia; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology*; D016865:Reassortant Viruses / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013552:Swine