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Detection of influenza A virus from live-bird market poultry swab samples in China by a pan-IAV, one-step reverse-transcription FRET-PCR

tetano

Editor, Senior Moderator
Sci Rep. 2016 Jul 22;6:30015. doi: 10.1038/srep30015.
[h=1]Detection of influenza A virus from live-bird market poultry swab samples in China by a pan-IAV, one-step reverse-transcription FRET-PCR.[/h] Luan L[SUP]1[/SUP], Sun Z[SUP]1[/SUP], Kaltenboeck B[SUP]2[/SUP], Huang K[SUP]1[/SUP], Li M[SUP]1[/SUP], Peng D[SUP]1[/SUP], Xu X[SUP]1[/SUP], Ye J[SUP]1[/SUP], Li J[SUP]1[/SUP], Guo W[SUP]1[/SUP], Wang C[SUP]1,[/SUP][SUP]2[/SUP].
[h=3]Author information[/h]

[h=3]Abstract[/h] The persistent public health threat of animal to human transmission of influenza A virus (IAV) has stimulated interest in rapid and accurate detection of all IAV subtypes in clinical specimens of animal origin. In this study, a new set of primers and probes was designed for one-step pan-IAV reverse-transcription fluorescence resonance energy transfer (FRET)-PCR. The detection limit of one-step pan-IAV RT FRET-PCR was 10 copies of the matrix gene per reaction, and proved to be equivalent or superior to virus isolation in detecting nine IAV subtypes. Application of the pan-IAV RT FRET-PCR to oral-pharyngeal and cloacal swab specimens collected from healthy poultry in 34 live bird markets in 24 provinces of China revealed that 9.2% of the animals (169/1,839) or 6.3% of their oral-pharyngeal or cloacal swabs (233/3,678) were positive for IAV, and 56.8% of IAV-positive samples were of the H9N2 subtype. Paralleling detection of IAV in H9N2-infected SPF chickens and chickens from LBM showed that pan-IAV FRET-PCR had a higher detection limit than virus isolation in eggs while the results by FRET-PCR and virus isolation overall matched. It is expected that this strategy can be useful for facile surveillance for IAV in clinical samples from a variety of sources.


PMID: 27445010 DOI: 10.1038/srep30015
[PubMed - in process] Free full text
 
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