Development and Evaluation of Multiplex Real-time RT-PCR Assays for Seasonal, Pandemic A/H1pdm09 and Avian A/H5 Influenza Viruses Detection

Jang-Hoon Choi; Mi-Seon Kim; Joo-Yeon Lee; Nam-Joo Lee; Donghyok Kwon; Min Gu Kang; Chun Kang

doi:10.1007/s12275-013-2452-y

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Research Support, Non-U.S. Gov't
Development and Evaluation of Multiplex Real-time RT-PCR Assays for Seasonal, Pandemic A/H1pdm09 and Avian A/H5 Influenza Viruses Detection: Jang-Hoon Choi , Mi-Seon Kim , Joo-Yeon Lee , Nam-Joo Lee , Donghyok Kwon , Min Gu Kang , Chun Kang; Journal of Microbiology 2013;51(2):252-257.
DOI: https://doi.org/10.1007/s12275-013-2452-y
Published online: April 27, 2013

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

Corresponding author: Chun Kang , Tel: +82-43-719-8190,

Received: 29 August 2012 • Accepted: 2 January 2013

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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 RTPCR 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.

Keywords: influenza;real-time RT-PCR;surveillance;diagnosis

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Development and Evaluation of Multiplex Real-time RT-PCR Assays for Seasonal, Pandemic A/H1pdm09 and Avian A/H5 Influenza Viruses Detection

J. Microbiol. 2013;51(2):252-257. Published online April 27, 2013

DOI: https://doi.org/10.1007/s12275-013-2452-y

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