title: Design of a single tube RT-PCR assay for the diagnosis of human infection with highly pathogenic influenza A(H5) viruses.
abstract: Concerns about emergence of a pandemic strain of influenza have been increasing. The strains of highly pathogenic influenza A(H5N1) currently circulating are considered among the most plausible candidates for giving rise to a pandemic strain. In this study the design and development of a RT-PCR assay specific for these highly pathogenic influenza A(H5) strains is presented. This is achieved in part by the design of a primer targeting the coding region for the protease cleavage site of the hemagglutinin, and another primer derived from a pan-hemagglutinin RT-PCR assay also presented in this study. It is shown that the HPAI A(H5) specific assay amplifies only the nucleic acids of highly pathogenic A(H5), with a high sensitivity.
authors.affiliation: Metabolism Research Program, Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada.
authors.forename: C
authors.initials: C
authors.lastname: Yea
authors.forename: D
authors.initials: D
authors.lastname: Adachi
authors.forename: G
authors.initials: G
authors.lastname: Johnson
authors.forename: E
authors.initials: E
authors.lastname: Nagy
authors.forename: F
authors.initials: F
authors.lastname: Gharabaghi
authors.forename: M
authors.initials: M
authors.lastname: Petric
authors.forename: S E
authors.initials: SE
authors.lastname: Richardson
authors.forename: R
authors.initials: R
authors.lastname: Tellier
mesh_terms: D000818:Animals; D001717:Birds; D017931:DNA Primers; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D005585:Influenza in Birds / Q000175:diagnosis / Q000821:virology; D007251:Influenza, Human / Q000175:diagnosis*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D012367:RNA, Viral