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

作者：Luan, Lu; Sun, Zhihao; Kaltenboeck, Bernhard; Huang, Ke; Li, Min; Peng, Daxin; Xu, Xiulong; Ye, Jianqiang; Li, Jing; Guo, Weina; Wang, Chengming^*

来源：Scientific Reports, 2016, 6(1): 30015.

DOI：10.1038/srep30015

• MULTIPLEX RT-PCR
• EASTERN CHINA
• A(H7N9) VIRUS
• ASSAY
• AMPLIFICATION
• TRANSMISSION
• SUBTYPES
• QPCR
• H7N9
• H5N1

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.

• 出版日期2016-7-22
• 单位扬州大学