title: Development and evaluation of multiplex real-time RT-PCR assays for seasonal, pandemic A/H1pdm09 and avian A/H5 influenza viruses detection.
abstract: Since the pandemic influenza A (H1N1) 2009 ((H1N1)pdm09) virus spread all over the world, the (H1N1)pdm09 virus has been circulating with seasonal influenza viruses. We developed rapid and sensitive one-step multiplex real-time RT-PCR assays (rRT-PCR) for simultaneous detection of influenza viruses currently circulating in humans, and the avian A/H5 virus. The detection limit of each assay was 4.8 to 1 copies per reaction and no cross-reactivity with other major respiratory pathogens was found. Analytical positive predictive value (PPV), negative predictive value (NPV) sensitivity and specificity were 100%, 94.1%, 93.7% and 100%, respectively. Clinical evaluation revealed that 1,976 (16.5%) of 11,963 throat swabs from patients with respiratory symptoms were confirmed as 1,651 (83.6%) A/H1pdm09, 308 (15.6%) A/H3 and 17 (0.8%) B virus during the 2010-2011 influenza season. Collectively, the multiplex rRT-PCR assays described here provide a practical tool for reliable implementation of influenza surveillance and diagnosis.
authors.affiliation: Division of Influenza Virus, Center for Infectious Disease, Korea National Institute of Health, Korea Center for Disease Control and Prevention, Osong 363-951, Republic of Korea.
authors.forename: Jang-Hoon
authors.initials: JH
authors.lastname: Choi
authors.forename: Mi-Seon
authors.initials: MS
authors.lastname: Kim
authors.forename: Joo-Yeon
authors.initials: JY
authors.lastname: Lee
authors.forename: Nam-Joo
authors.initials: NJ
authors.lastname: Lee
authors.forename: Donghyok
authors.initials: D
authors.lastname: Kwon
authors.forename: Min Gu
authors.initials: MG
authors.lastname: Kang
authors.forename: Chun
authors.initials: C
authors.lastname: Kang
mesh_terms: D017931:DNA Primers / Q000235:genetics; D005260:Female; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D053124:Influenza A Virus, H5N1 Subtype / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology*; D058873:Pandemics; D056910:Republic of Korea / Q000453:epidemiology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012621:Seasons; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers