title: Development of a fluorescent probe-based recombinase polymerase amplification assay for rapid detection of Orf virus.
abstract: BACKGROUND
  Orf virus (ORFV) is the causative agent of Orf (also known as contagious ecthyma or contagious papular dermatitis), a severe infectious skin disease in goats, sheep and other ruminants. The rapid detection of ORFV is of great importance in disease control and highly needed. A isothermal molecular diagnostic approach, termed recombinase polymerase amplification (RPA), is considered as an novel and rapid alternative techonology to PCR assay.
  
  
  RESULTS
  In the present study, a novel fluorescent probe based on RPA assay (ORFV exo RPA assay) was developed. The developed ORFV exo RPA assay was capable of as low as 100 copies of ORFV DNA /reaction and was highly specific, with no cross-reaction with closely related viruses (capripox virus, foot-and-mouth disease virus or peste des petits ruminants virus). Further assessment with clinical samples showed that the developed ORFV exo RPA assay has good correlation with qPCR assays for detection of ORFV.
  
  
  CONCLUSIONS
  These results suggest that the developed ORFV exo RPA assay is suitable for rapid detection of ORFV.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China. yangyang8706@163.com.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China. qinxiaodong@caas.cn.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Qin
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China. wangguangxiang@caas.cn.
authors.forename: Guangxiang
authors.initials: G
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China. zhang599@hotmail.co.uk.|The Medical School, University College London, Gower Street, London, WC1E 6BT, UK. zhang599@hotmail.co.uk.
authors.forename: Yuen
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China. shangyoujun@caas.cn.
authors.forename: Youjun
authors.initials: Y
authors.lastname: Shang
authors.affiliation: 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.
authors.forename: Zhidong
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D000818:Animals; D003429:Cross Reactions; D004474:Ecthyma, Contagious / Q000175:diagnosis* / Q000821:virology; D005456:Fluorescent Dyes; D006041:Goats; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009923:Orf virus / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction; D045522:Recombinases / Q000378:metabolism; D012680:Sensitivity and Specificity; D012756:Sheep; D013997:Time Factors
chemical_list: D005456:Fluorescent Dyes; D045522:Recombinases