title: Rapid detection of highly pathogenic porcine reproductive and respiratory syndrome virus by a fluorescent probe-based isothermal recombinase polymerase amplification assay.
abstract: A novel fluorescent probe-based real-time reverse transcription recombinase polymerase amplification (real-time RT-RPA) assay was developed for rapid detection of highly pathogenic type 2 porcine reproductive and respiratory syndrome virus (HP-PRRSV). The sensitivity analysis showed that the detection limit of RPA was 70 copies of HP-PRRSV RNA/reaction. The real-time RT-RPA highly specific amplified HP-PRRSV with no cross-reaction with classic PRRSV, classic swine fever virus, pseudorabies virus, and foot-and-mouth disease virus. Assessment with 125 clinical samples showed that the developed real-time RT-RPA assay was well correlated with real-time RT-qPCR assays for detection of HP-PRRSV. These results suggest that the developed real-time RT-RPA assay is suitable for rapid detection of HP-PRRSV.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Qin
authors.affiliation: Shandong New Hope Liuhe Co., Ltd., Qingdao, 266000, Shandong, China.
authors.forename: Yingjun
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Shandong New Hope Liuhe Co., Ltd., Qingdao, 266000, Shandong, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China. zhangzhidong@caas.cn.
authors.forename: Zhidong
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D000818:Animals; D005456:Fluorescent Dyes; D005814:Genes, Viral; D021141:Nucleic Acid Amplification Techniques; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis* / Q000821:virology*; D019316:Porcine respiratory and reproductive syndrome virus / Q000145:classification / Q000235:genetics*; D012367:RNA, Viral; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013552:Swine
chemical_list: D005456:Fluorescent Dyes; D012367:RNA, Viral
keywords: HP-PRRSV; Molecular diagnosis; RT-qPCR; Real-time RT-RPA; Recombinase polymerase amplification