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

tetano

Editor, Senior Moderator
Viruses. 2020 Jan 15;12(1). pii: E100. doi: 10.3390/v12010100. [h=1]Development of a Multiplex RT-qPCR for the Detection of Different Clades of Avian Influenza in Poultry.[/h]
Le TB[SUP]1,[/SUP][SUP]2[/SUP], Kim HK[SUP]3[/SUP], Na W[SUP]4[/SUP], Le VP[SUP]5[/SUP], Song MS[SUP]6[/SUP], Song D[SUP]7[/SUP], Jeong DG[SUP]1,[/SUP][SUP]2[/SUP], Yoon SW[SUP]1,[/SUP][SUP]2[/SUP].
[h=3]Author information[/h]

[h=3]Abstract[/h] 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.


[h=4]KEYWORDS:[/h] Clade; H5Nx; HPAI; field samples; simultaneous detection

PMID: 31952218 DOI: 10.3390/v12010100
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