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Open AccessArticle

Development of a Multiplex RT-qPCR for the Detection of Different Clades of Avian Influenza in Poultry

1
Infectious Diseases Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Korea
2
Bio-Analytical Science Division, University of Science and Technology, Daejeon 34113, Korea
3
Department of Microbiology, Chungbuk National University, Cheongju 28644, Korea
4
College of Veterinary Medicine, Chonnam National University, Gwangju 61186, Korea
5
College of Veterinary Medicine, Vietnam National University of Agriculture, Hanoi 100000, Vietnam
6
College of Medicine, Chungbuk National University, Cheongju 28644, Korea
7
College of Pharmacy, Korea University, Sejong City 30019, Korea
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Viruses 2020, 12(1), 100; https://doi.org/10.3390/v12010100
Received: 6 December 2019 / Revised: 9 January 2020 / Accepted: 13 January 2020 / Published: 15 January 2020
(This article belongs to the Section Animal Viruses)
Since the initial detection of H5N1, a highly pathogenic avian influenza (HPAI) virus, in 1996 in China, numerous HPAI H5 lineages have been classified, and they continue to pose a threat to animal and human health. In this study, we developed a novel primer/probe set that can be employed to simultaneously detect pan-H5 HPAI and two clades, 2.3.2.1 and 2.3.4.4, of H5Nx viruses using reverse transcription quantitative polymerase chain reaction (RT-qPCR). The sensitivity and specificity of these primer sets and probes were confirmed with a number of different subtypes of influenza virus and the H5-HA gene plasmid DNA. In particular, the multiplex RT-qPCR assay was successfully applied to the simultaneous detection of H5 HPAI and different virus clades in clinical field samples from a poultry farm. Therefore, this multiplex assay and a novel detection primer set and probes will be useful for the laboratory diagnosis and epidemiological field studies of different circulating H5 HPAI virus clades in poultry and migratory wild birds. View Full-Text
Keywords: HPAI; H5Nx; Clade; simultaneous detection; field samples HPAI; H5Nx; Clade; simultaneous detection; field samples
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MDPI and ACS Style

Le, T.B.; Kim, H.K.; Na, W.; Le, V.P.; Song, M.-S.; Song, D.; Jeong, D.G.; Yoon, S.-W. Development of a Multiplex RT-qPCR for the Detection of Different Clades of Avian Influenza in Poultry. Viruses 2020, 12, 100.

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