Development of real-time and lateral flow strip reverse transcription recombinase polymerase Amplification assays for rapid detection of peste des petits ruminants virus.
Peste des petits ruminants (PPR) is an economically important, Office International des Epizooties (OIE) notifiable, transboundary viral disease of small ruminants such as sheep and goat. PPR virus (PPRV), a negative-sense single-stranded RNA virus, is the causal agent of PPR. Therefore, sensitive, specific and rapid diagnostic assay for the detection of PPRV are necessary to accurately and promptly diagnose suspected case of PPR.



In this study, reverse transcription recombinase polymerase amplification assays using real-time fluorescent detection (real-time RT-RPA assay) and lateral flow strip detection (LFS RT-RPA assay) were developed targeting the N gene of PPRV.



The sensitivity of the developed real-time RT-RPA assay was as low as 100 copies per reaction within 7 min at 40 °C with 95% reliability; while the sensitivity of the developed LFS RT-RPA assay was as low as 150 copies per reaction at 39 °C in less than 25 min. In both assays, there were no cross-reactions with sheep and goat pox viruses, foot-and-mouth disease virus and Orf virus.



These features make RPA assay promising candidates either in field use or as a point of care diagnostic technique.
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
Yang
Y
Yang
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
Xiaodong
X
Qin
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
Yiming
Y
Song
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
Wei
W
Zhang
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
Gaowei
G
Hu
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
Yongxi
Y
Dou
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
Yanmin
Y
Li
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.
Zhidong
Z
Zhang
D000818:Animals; D003429:Cross Reactions; D005456:Fluorescent Dyes; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics / Q000302:isolation & purification; D015511:Goat Diseases / Q000175:diagnosis / Q000821:virology; D006041:Goats / Q000821:virology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods* / Q000662:veterinary*; D009923:Orf virus / Q000235:genetics / Q000302:isolation & purification; D029021:Peste-des-Petits-Ruminants / Q000175:diagnosis* / Q000821:virology; D018107:Peste-des-petits-ruminants virus / Q000235:genetics* / Q000302:isolation & purification*; D012367:RNA, Viral / Q000302:isolation & purification; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D012756:Sheep / Q000821:virology; D012757:Sheep Diseases / Q000175:diagnosis / Q000821:virology
D005456:Fluorescent Dyes; D012367:RNA, Viral
Lateral flow strip; PPRV; RT-RPA; Reverse transcription recombinase polymerase amplification assay; Small ruminants