title: Comparison and evaluation of conventional RT-PCR, SYBR green I and TaqMan real-time RT-PCR assays for the detection of porcine epidemic diarrhea virus.
abstract: Porcine epidemic diarrhea (PED) caused by porcine epidemic diarrhea virus (PEDV) is a highly contagious intestinal disease, resulting in substantial economic losses to the swine industry worldwide. In this study, three assays, namely a conventional reverse transcription-polymerase chain reaction (RT-PCR), a SYBR Green I real-time RT-PCR and a TaqMan real-time RT-PCR targeting the highly conserved M gene of PEDV, were developed and evaluated. Then, the analytical specificity, sensitivity and reproducibility of these assays were determined and compared. The TaqMan real-time RT-PCR was 100-fold and 10,000-fold more sensitive than that of the SYBR Green I real-time RT-PCR and the conventional RT-PCR, respectively. The analytical sensitivity of TaqMan real-time RT-PCR was 10 copies/μl of target gene and no cross amplification with other viruses tested was observed. With the features of high specificity, sensitivity, and reproducibility, the TaqMan real-time RT-PCR established in this study could be a useful tool for clinical diagnosis, epidemiological surveys and outbreak investigations of PED.
authors.affiliation: Department of Preventive Veterinary Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, 330045 China. Electronic address: 13732944385@163.com.
authors.forename: Xinrong
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of Preventive Veterinary Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, 330045 China. Electronic address: zhangtiansheng0512@163.com.
authors.forename: Tiansheng
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Department of Preventive Veterinary Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, 330045 China. Electronic address: sdpjxau@hotmail.com.
authors.forename: Deping
authors.initials: D
authors.lastname: Song
authors.affiliation: Department of Preventive Veterinary Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, 330045 China. Electronic address: 402983803@qq.com.
authors.forename: Tao
authors.initials: T
authors.lastname: Huang
authors.affiliation: Department of Preventive Veterinary Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, 330045 China. Electronic address: jxaudkypq@hotmail.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Peng
authors.affiliation: Department of Preventive Veterinary Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, 330045 China. Electronic address: 185704818@qq.com.
authors.forename: Yanjun
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Preventive Veterinary Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, 330045 China. Electronic address: lianqi19920826@163.com.
authors.forename: Anqi
authors.initials: A
authors.lastname: Li
authors.affiliation: Department of Preventive Veterinary Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, 330045 China. Electronic address: zfanfan816@qq.com.
authors.forename: Fanfan
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Department of Preventive Veterinary Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, 330045 China. Electronic address: lbls2005@sina.com.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Wu
authors.affiliation: Department of Preventive Veterinary Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, 330045 China. Electronic address: yy6157832@163.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Ye
authors.affiliation: Department of Preventive Veterinary Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, 330045 China. Electronic address: tang53ster@gmail.com.
authors.forename: Yuxin
authors.initials: Y
authors.lastname: Tang
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D018352:Coronavirus Infections / Q000175:diagnosis* / Q000235:genetics / Q000821:virology; D003959:Diamines; D009930:Organic Chemicals / Q000737:chemistry; D053485:Porcine epidemic diarrhea virus / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013552:Swine / Q000821:virology; D013553:Swine Diseases / Q000175:diagnosis* / Q000235:genetics / Q000821:virology
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I
keywords: PEDV; SYBR green I real-time RT-PCR; Sensitivity; Specificity; TaqMan real-time RT-PCR