title: Development of an isothermoal amplification-based assay for rapid visual detection of an Orf virus.
abstract: BACKGROUND
  Orf virus (ORFV) is the causative agent of a severe infectious skin disease (also known as contagious ecthyma) in goats, sheep and other small ruminants. Importantly, ORFV also infect humans which causes a public health concern in the context of changing environment and increase in human populations. The rapid detection is critical in effective control of the disease and urgently needed.
  
  
  RESULTS
  A novel "point of care" molecular amplification assay for rapid visual detection of ORFV was developed based on isothermoal recombinase polymerase amplification (RPA) technology in combination with a simpler lateral flow immunoassay strip (ORFV RPA- LFD assay). The developed ORFV RPA- LFD assay was able to detect ORFV in less than 25 min. This assay was highly sensitive, with detection limit of as low as 80 copies/reaction, and highly specific, with no cross-reactions with capripox virus, foot-and-mouth disease virus and peste des petits ruminants virus. Furthermore, the ORFV RPA- LFD assay has good correlation with qPCR assay for detection of ORFV present in clinical samples.
  
  
  CONCLUSIONS
  The developed ORFV RPA-LFD assay was a sensitive and specific method for rapid detection of ORFV, and has great potential as an onsite molecular diagnostic tool in control of Orf.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Lanzhou, Gansu, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Lanzhou, Gansu, China.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Qin
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Lanzhou, Gansu, China.
authors.forename: Guangxiang
authors.initials: G
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Lanzhou, Gansu, China.|School of Life Science, Lanzhou University, Lanzhou, Gansu, China.
authors.forename: Jiaxin
authors.initials: J
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Lanzhou, Gansu, China.
authors.forename: Youjun
authors.initials: Y
authors.lastname: Shang
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Lanzhou, Gansu, China. zhangzhidong@caas.cn.
authors.forename: Zhidong
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D000818:Animals; D004279:DNA, Viral; D004474:Ecthyma, Contagious / Q000175:diagnosis* / Q000821:virology*; D005814:Genes, Viral; D006041:Goats; D021141:Nucleic Acid Amplification Techniques; D009923:Orf virus / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral
keywords: Isothermal nucleic acid amplification; Lateral flow; Orf virus; RPA; Rapid test; Recombinase polymerase amplification