title: Detection of influenza A virus from live-bird market poultry swab samples in China by a pan-IAV, one-step reverse-transcription FRET-PCR.
abstract: 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.
authors.affiliation: Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu Province, P. R. China.
authors.forename: Lu
authors.initials: L
authors.lastname: Luan
authors.affiliation: Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu Province, P. R. China.
authors.forename: Zhihao
authors.initials: Z
authors.lastname: Sun
authors.affiliation: College of Veterinary Medicine, Auburn University, Auburn, Alabama, USA.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Kaltenboeck
authors.affiliation: Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu Province, P. R. China.
authors.forename: Ke
authors.initials: K
authors.lastname: Huang
authors.affiliation: Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu Province, P. R. China.
authors.forename: Min
authors.initials: M
authors.lastname: Li
authors.affiliation: Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu Province, P. R. China.
authors.forename: Daxin
authors.initials: D
authors.lastname: Peng
authors.affiliation: Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu Province, P. R. China.
authors.forename: Xiulong
authors.initials: X
authors.lastname: Xu
authors.affiliation: Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu Province, P. R. China.
authors.forename: Jianqiang
authors.initials: J
authors.lastname: Ye
authors.affiliation: Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu Province, P. R. China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu Province, P. R. China.
authors.forename: Weina
authors.initials: W
authors.lastname: Guo
authors.affiliation: Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu Province, P. R. China.|College of Veterinary Medicine, Auburn University, Auburn, Alabama, USA.
authors.forename: Chengming
authors.initials: C
authors.lastname: Wang
mesh_terms: D000818:Animals; D002645:Chickens; D002681:China; D002988:Cloaca / Q000821:virology; D017931:DNA Primers / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D009980:Influenza A virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009055:Mouth / Q000821:virology; D015345:Oligonucleotide Probes / Q000235:genetics; D010614:Pharynx / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D015345:Oligonucleotide Probes