title: Rapid and sensitive detection of canine distemper virus by one-tube reverse transcription-insulated isothermal polymerase chain reaction.
abstract: BACKGROUND
  Canine distemper virus (CDV) has been associated with outbreaks of canine infectious respiratory disease in shelters and boarding kennel environments. POCKITTM Nucleic Acid Analyzer is a field-deployable device capable of generating automatically interpreted insulated isothermal polymerase chain reaction (iiPCR) results from extracted nucleic acid within one hour. In this study, reverse transcription iiPCR (RT-iiPCR) was developed to facilitate point-of-need diagnosis of CDV infection.
  
  
  RESULTS
  Analytical sensitivity (limit of detection 95%) of the established CDV RT-iiPCR was about 11 copies of in vitro transcribed RNA per reaction. CDV RT-iiPCR generated positive signals from CDV, but not Bordetella bronchiseptica, canine parvovirus, canine herpesvirus, canine adenovirus 2, canine influenza virus (subtype H3N8), canine parainfluenza virus, and canine respiratory coronavirus. To evaluate accuracy of the established reaction in canine distemper clinical diagnosis, 110 specimens from dogs, raccoons, and foxes suspected with CDV infection were tested simultaneously by CDV RT-iiPCR and real-time RT-PCR. CDV RT-iiPCR demonstrated excellent sensitivity (100%) and specificity (100%), compared to real-time RT-PCR.
  
  
  CONCLUSIONS
  The results indicated an excellent correlation between RT-iiPCR and a reference real time RT-PCR method. Working in a lyophilized format, the established method has great potential to be used for point-of-care diagnosis of canine distemper in animals, especially in resource-limited facilities.
authors.forename: Rebecca P
authors.initials: RP
authors.lastname: Wilkes
authors.forename: Yun-Long
authors.initials: YL
authors.lastname: Tsai
authors.forename: Pei-Yu
authors.initials: PY
authors.lastname: Lee
authors.forename: Fu-Chun
authors.initials: FC
authors.lastname: Lee
authors.forename: Hsiao-Fen Grace
authors.initials: HF
authors.lastname: Chang
authors.forename: Hwa-Tang Thomas
authors.initials: HT
authors.lastname: Wang
mesh_terms: D000818:Animals; D004216:Distemper / Q000175:diagnosis / Q000821:virology*; D004217:Distemper Virus, Canine / Q000302:isolation & purification*; D004285:Dogs; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012367:RNA, Viral / Q000302:isolation & purification; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D012367:RNA, Viral