title: Rapid and real-time detection of Porcine Sapelovirus by reverse transcription loop-mediated isothermal amplification assay.
abstract: The present study describes the development and validation of a one-step, single-tube, and real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) detecting Porcine Sapelovirus. RT-LAMP characterized by one strand displacement reaction with the specific stem-loop structure and Bst DNA polymerase could be finished in 60 min under isothermal condition at 63 °C. RT-LAMP assay showed higher sensitivity with 10(1) copies/μL than RT-PCR for the detection of Sapelovirus. The specificity of RT-LAMP assay was validated by the absence of any cross-reaction with other closely related virus in Picornaviridae group and other common virus causing porcine diarrhea. 7 positive Sapelovirus infection out of 63 fecal samples were identified using RT-LAMP, while 5 positive samples were determined by a conventional RT-PCR. A cost-effective method for Saplovirus detection with high sensitivity and specificity was developed and evaluated.
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, China. Electronic address: wcy_an@sina.cn.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Wang
authors.affiliation: Animal Disease Control Center of Minhang District, 1633 Humin Road, Shanghai, China. Electronic address: mhqsys@126.com.
authors.forename: Dayi
authors.initials: D
authors.lastname: Yu
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, China. Electronic address: lcui@sjtu.edu.cn.
authors.forename: Li
authors.initials: L
authors.lastname: Cui
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, China. Electronic address: hxg@sjtu.edu.cn.
authors.forename: Xiuguo
authors.initials: X
authors.lastname: Hua
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, China. Electronic address: ycl@sjtu.edu.cn.
authors.forename: Congli
authors.initials: C
authors.lastname: Yuan
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, China. Electronic address: sunhuan198419@163.com.
authors.forename: Huan
authors.initials: H
authors.lastname: Sun
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, China. Electronic address: 313081543@qq.com.
authors.forename: Yuxiao
authors.initials: Y
authors.lastname: Liu
mesh_terms: D000818:Animals; D003365:Costs and Cost Analysis; D005243:Feces / Q000821:virology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010849:Picornaviridae / Q000302:isolation & purification*; D010850:Picornaviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology*; D013696:Temperature; D013997:Time Factors
keywords: Detection; Picornaviridae; Porcine Sapelovirus; RT-LAMP