title: Development of a novel immuno-PCR for detection of avian leukosis virus.
abstract: Avian leukosis virus (ALV) is an important pathogen for various neoplasms, including lymphoid, myeloid, and erythroid neoplasms, and it causes significant economic loss in the poultry industry. Several efficient methods for the detection of ALV have been reported. However, these previously developed approaches are based on either PCR or immunoassays. Here, we used a proximity ligation technique and combined PCR with the immunoassay to develop a novel immuno-PCR (Im-PCR) approach for the detection of ALV. Our data showed that the Im-PCR had high specificity and sensitivity to ALV. The Im-PCR method selectively reacted to ALV but not to the other avian viruses tested. The limit of detection of Im-PCR could reach 0.5 TCID50. Moreover, the results of Im-PCR were in agreement with results from commercial ELISA when the clinical cloaca samples were used for ALV detection. The present results demonstrate that the novel Im-PCR method can be efficiently applied to detect ALV in a clinical setting. Our data also highlight that Im-PCR may have promising applications in the diagnosis of pathogens.
authors.affiliation: Ministry of Education Key Laboratory for Avian Preventive Medicine, and Key Laboratory of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, PR China.
authors.forename: Quan
authors.initials: Q
authors.lastname: Xie
authors.affiliation: Sinopharm Yangzhou VAC Biological Engineering Co., Ltd., Yangzhou, Jiangsu, PR China.
authors.forename: Jianjun
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Ministry of Education Key Laboratory for Avian Preventive Medicine, and Key Laboratory of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, PR China.
authors.forename: Hongxia
authors.initials: H
authors.lastname: Shao
authors.affiliation: Ministry of Education Key Laboratory for Avian Preventive Medicine, and Key Laboratory of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, PR China.
authors.forename: Zhimin
authors.initials: Z
authors.lastname: Wan
authors.affiliation: Ministry of Education Key Laboratory for Avian Preventive Medicine, and Key Laboratory of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, PR China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Tian
authors.affiliation: Institute of Genomics and Multiscale Biology, Icahn School of Medicine at Mount Sinai, NY, USA.
authors.forename: Jialiang
authors.initials: J
authors.lastname: Yang
authors.affiliation: Institute of Animal Health Inspection in Wujiang, Suzhou, Jiangsu, PR China.
authors.forename: Mayun
authors.initials: M
authors.lastname: Pang
authors.affiliation: Ministry of Education Key Laboratory for Avian Preventive Medicine, and Key Laboratory of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, PR China.
authors.forename: Kun
authors.initials: K
authors.lastname: Qian
authors.affiliation: Ministry of Education Key Laboratory for Avian Preventive Medicine, and Key Laboratory of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Gao
authors.affiliation: Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, PR China.
authors.forename: Chengming
authors.initials: C
authors.lastname: Wang
authors.affiliation: Ministry of Education Key Laboratory for Avian Preventive Medicine, and Key Laboratory of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, PR China. Electronic address: aijian@yzu.edu.cn.
authors.forename: Aijian
authors.initials: A
authors.lastname: Qin
authors.affiliation: Ministry of Education Key Laboratory for Avian Preventive Medicine, and Key Laboratory of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, PR China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, PR China. Electronic address: jqye@yzu.edu.cn.
authors.forename: Jianqiang
authors.initials: J
authors.lastname: Ye
mesh_terms: D000818:Animals; D001353:Avian Leukosis / Q000175:diagnosis* / Q000821:virology; D001354:Avian Leukosis Virus / Q000302:isolation & purification*; D002988:Cloaca / Q000821:virology*; D007118:Immunoassay / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011200:Poultry; D012680:Sensitivity and Specificity
keywords: Avian leukosis virus; Detection; Immuno-PCR; Proximity ligation assay