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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

tetano

Editor, Senior Moderator
J Microbiol. 2013 Apr;51(2):252-7. doi: 10.1007/s12275-013-2452-y. Epub 2013 Apr 27.
Development and evaluation of multiplex real-time RT-PCR assays for seasonal, pandemic A/H1pdm09 and avian A/H5 influenza viruses detection.
Choi JH, Kim MS, Lee JY, Lee NJ, Kwon D, Kang MG, Kang C.
Source

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.
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.

PMID:
23625229
[PubMed - in process]

http://www.ncbi.nlm.nih.gov/pubmed/23625229
 
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