Abstract:
Sheeppox virus (SPPV), goatpox virus (GTPV) and lumpy skin disease virus (LSDV) belong to the genus Capripoxvirus (CaPV), and are important pathogens of sheep, goat and cattle, respectively. Rapid and reliable detection of CaPV is critical to prevent its spread and promote its eradication. This study aimed to develop the recombinase polymerase amplification (RPA) assays combined with real-time fluorescence (real-time RPA) and naked-eye visible lateral flow strip (LFS RPA) for rapid detection of CaPV. Both developed RPA assays worked well at 39 °C within 20 min. They were highly specific for the detection of GTPV, SPPV and LSDV, while no cross-reactivity was observed for other non-targeted pathogens and genomic DNA of goat, sheep and cattle. The limit of detection for real-time RPA and LFS RPA were 1.0 × 102 and 1.0 × 101 copies per reaction, respectively. In the artificially contaminated samples with GTPV, the detection results of RPA assays were consistent with those of real-time PCR. For 15 clinical samples, LSDV was detected by real-time RPA, LFS RPA and real-time PCR in 13, 15 and 15 samples, respectively. The developed RPA assays were specific, sensitive, and user-friendly for the rapid detection of CaPV, and could be a better alternative method applied in low-resources settings.
摘要:
羊痘病毒(SPPV)山羊痘病毒(GTPV)和块状皮肤病病毒(LSDV)属于羊痘病毒属(CaPV),是绵羊的重要病原体,山羊和牛,分别。快速可靠地检测CaPV对于防止其传播和促进其根除至关重要。本研究旨在开发重组酶聚合酶扩增(RPA)结合实时荧光(实时RPA)和肉眼可见侧流带(LFSRPA)的快速检测CaPV的方法。两种开发的RPA测定在39°C下在20min内工作良好。它们对GTPV的检测具有高度特异性,SPPV和LSDV,虽然没有观察到其他非靶向病原体和山羊基因组DNA的交叉反应,羊和牛。实时RPA和LFSRPA的检测限为每个反应1.0×102和1.0×101个拷贝,分别。在用GTPV人工污染的样品中,RPA检测结果与real-timePCR检测结果一致。对于15个临床样本,通过实时RPA检测LSDV,13、15和15个样品的LFSRPA和实时PCR,分别。开发的RPA测定法是特异性的,敏感,和用户友好的快速检测CaPV,并且可能是在低资源环境中应用的更好的替代方法。