Development of real-time and lateral flow dipstick recombinase polymerase amplification assays for rapid detection of goatpox virus and sheeppox virus

Virol J. 2017 Jul 17;14(1):131. doi: 10.1186/s12985-017-0792-7.

Abstract

Background: Goatpox 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.

Methods: Recombinase 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.

Results: The 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.

Conclusions: This study provided a highly time-efficient and simple alternative for rapid detection of GTPV and SPPV.

Keywords: CaPV RPA LFD; CaPV real-time RPA; Goatpox virus; Recombinase polymerase amplification; Sheeppox virus.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Capripoxvirus / isolation & purification*
  • DNA, Viral / analysis*
  • DNA, Viral / genetics
  • Goat Diseases / diagnosis*
  • Goat Diseases / virology
  • Goats
  • Molecular Diagnostic Techniques / methods*
  • Polymerase Chain Reaction / methods*
  • Poxviridae Infections / diagnosis
  • Poxviridae Infections / veterinary*
  • Poxviridae Infections / virology
  • Sensitivity and Specificity
  • Sheep
  • Sheep Diseases / diagnosis*
  • Sheep Diseases / virology
  • Temperature
  • Time Factors
  • Veterinary Medicine / methods

Substances

  • DNA, Viral