title: Reverse Transcription Cross-Priming Amplification-Nucleic Acid Test Strip for Rapid Detection of Porcine Epidemic Diarrhea Virus.
abstract: Porcine epidemic diarrhea virus (PEDV) is a highly transmissible coronavirus that causes a severe enteric disease particularly in neonatal piglets. In this study, a rapid method for detecting PEDV was developed based on cross-priming amplification and nucleic acid test strip(CPA-NATS). Five primers specific for the N gene sequence of PEDV were used for the cross-priming amplification. Detection of amplification products based on labeled probe primers was conducted with strip binding antibody of labeled markers. The CPA method was evaluated and compared with a PCR method. The reverse transcription CPA system was further optimized for detecting PEDV RNA in clinical specimens. Results showed that the method was highly specific for the detection of PEDV, and had the same sensitivity as PCR, with detection limit of 10(-6) diluted plasmid containing the target gene of PEDV. It was also successfully applied to detecting PEDV in clinical specimens. The reverse transcription CPA-NATS detection system established in this study offers a specific, sensitive, rapid, and simple detection tool for screening PEDV, which can contribute to strategies in the effective control of PEDV in swine.
authors.affiliation: State Key Laboratory of Special Economic Animal Molecular Biology, Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, No. 4899 Juye Avenue, Jingyue Economic and Technological Development Zone, Changchun, Jilin, 130112, People's Republic of China.
authors.forename: Feng-Xue
authors.initials: FX
authors.lastname: Wang
authors.affiliation: (Sino-USA) SiChuan Nabii Bio-Tech Co., Ltd., Chengdu, SiChuan, 610041, People's Republic of China.
authors.forename: Dan-Yi
authors.initials: DY
authors.lastname: Yuan
authors.affiliation: Ustar Biotechnologies (Hangzhou), Ltd., Hangzhou, Zhejiang, 310012, People's Republic of China.
authors.forename: Ya-Nan
authors.initials: YN
authors.lastname: Jin
authors.affiliation: Ustar Biotechnologies (Hangzhou), Ltd., Hangzhou, Zhejiang, 310012, People's Republic of China.
authors.forename: Lin
authors.initials: L
authors.lastname: Hu
authors.affiliation: (Sino-USA) SiChuan Nabii Bio-Tech Co., Ltd., Chengdu, SiChuan, 610041, People's Republic of China.
authors.forename: Zhi-Yong
authors.initials: ZY
authors.lastname: Sun
authors.affiliation: (Sino-USA) SiChuan Nabii Bio-Tech Co., Ltd., Chengdu, SiChuan, 610041, People's Republic of China.
authors.forename: Qian
authors.initials: Q
authors.lastname: He
authors.affiliation: Veterinary Research Institute, Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences, Huhhot, Inner Mongolia, 010031, People's Republic of China.
authors.forename: Shi-Hua
authors.initials: SH
authors.lastname: Zhao
authors.affiliation: Veterinary Research Institute, Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences, Huhhot, Inner Mongolia, 010031, People's Republic of China.
authors.forename: Shu-Bai
authors.initials: SB
authors.lastname: Zhan
authors.affiliation: State Key Laboratory of Special Economic Animal Molecular Biology, Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, No. 4899 Juye Avenue, Jingyue Economic and Technological Development Zone, Changchun, Jilin, 130112, People's Republic of China.|Veterinary Research Institute, Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences, Huhhot, Inner Mongolia, 010031, People's Republic of China.
authors.forename: Yong-Jun
authors.initials: YJ
authors.lastname: Wen
mesh_terms: D000368:Aged; D000818:Animals; D017931:DNA Primers; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D053485:Porcine epidemic diarrhea virus / Q000235:genetics / Q000302:isolation & purification*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology
chemical_list: D017931:DNA Primers