Abstract
BackgroundGoatpox virus (GTPV) and sheeppox virus (SPPV), which belong to the Capripoxvirus (CaPV), are economically important pathogens of small ruminants. Therefore, a sensitive, specific and rapid diagnostic assay for detection of GTPV and SPPV is necessary to accurately and promptly control these diseases.MethodsRecombinase polymerase amplification (RPA) assays combined with a real-time fluorescent detection (real-time RPA assay) and lateral flow dipstick (RPA LFD assay) were developed targeting the CaPV G-protein-coupled chemokine receptor (GPCR) gene, respectively.ResultsThe sensitivity of both CaPV real-time RPA assay and CaPV RPA LFD assay were 3 × 102 copies per reaction within 20 min at 38 °C. Both assays were highly specific for CaPV, with no cross-reactions with peste des petits ruminants virus, foot-and-mouth disease virus and Orf virus. The evaluation of the performance of these two assays with clinical sample (n = 107) showed that the CaPV real-time RPA assay and CaPV RPA LFD assay were able to specially detect SPPV or GTPV present in samples of ovine in liver, lung, kidney, spleen, skin and blood.ConclusionsThis study provided a highly time-efficient and simple alternative for rapid detection of GTPV and SPPV.
Highlights
Goatpox virus (GTPV) and sheeppox virus (SPPV), which belong to the Capripoxvirus (CaPV), are economically important pathogens of small ruminants
The results showed that both assays could detect viral DNA present in all the samples (Additional file 1: Table S1), and good correlation was found between threshold time (y axis) of CaPV real-time Recombinase polymerase amplification (RPA) assay and cycle threshold (CT) (x axis) of CaPV real-time Real-time quantitative polymerase chain reaction (PCR) (qPCR) assay (R squared 0.86, Fig. 3)
In evaluation of the specificity of CaPV real-time RPA assay, consistent positive signal was only observed for GTPV strains (GTPV AV40, GTPV AV41, GTPV GS-V1) and SPPV strains (SPPV Gulang2009, SPPV Jingtai2011, SPPV Hubei), and no cross detection was observed with other viruses which can infect sheep and goats, including foot-and-mouth disease virus (FMDV), Orf virus (ORFV) and peste des petits ruminants virus (PPRV) (Table 2)
Summary
Goatpox virus (GTPV) and sheeppox virus (SPPV), which belong to the Capripoxvirus (CaPV), are economically important pathogens of small ruminants. Sheeppox virus (SPPV) and goatpox virus (GTPV) belong to the genus Capripoxvirus (CaPV), subfamily Chordopoxvirinae, family Poxviridae, and cause serious pox diseases of domesticated small ruminants [1, 2]. For accurately and promptly controlling any outbreak, the foremost requirement is the sensitive, specific and rapid tool for detection of the causative agents [5, 6]. Various methods such as virus isolation, serology tests and Recombinase polymerase amplification (RPA) is a novel isothermal alternative to PCR, which can amplify detectable amount of DNA in 20 min or less with simple instrumentation.
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