Journal Article

Rapid and sensitive detection of canine distemper virus by one-tube reverse transcription-insulated isothermal polymerase chain reaction

Lee, Pei-Yu, Wang, Hwa-Tang Thomas, Tsai, Yun-Long, Lee, Fu-Chun, Chang, Hsiao-Fen Grace, Wilkes, Rebecca P

2014

Overview

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.

• Publisher: Springer-Verlag,BioMed Central,BioMed Central Ltd,Springer Nature B.V
• Subject: Acids / Animals / Bordetella bronchiseptica / Canid alphaherpesvirus 1 / Canid herpesvirus 1 / Canine adenovirus 2 / Canine adenovirus A / canine distemper / Canine distemper virus / Canine herpesvirus / Canine mastadenovirus A / Canine morbillivirus / Canine parvovirus / Carnivore protoparvovirus 1 / Coronavirinae / Coronavirus / Data analysis / Data processing / detection limit / Disease / Distemper - diagnosis / Distemper - virology / Distemper Virus, Canine - isolation & purification / Dogs / Feline panleukopenia virus / foxes / freeze drying / Health aspects / Infections / Influenza A virus / Influenza virus / Influenza viruses / Medical research / Medicine / Medicine & Public Health / Methodology / Methodology Article / Methods / Noise / Orthocoronavirinae / Parainfluenza virus / Polymerase chain reaction / Polymerase Chain Reaction - methods / Polymerase Chain Reaction - veterinary / Procyon lotor / Reproducibility of Results / reverse transcriptase polymerase chain reaction / reverse transcription / RNA / RNA, Viral - isolation & purification / Sensitivity and Specificity / Time Factors / Transgenics / Veterinary colleges / Veterinary medicine / Veterinary Medicine/Veterinary Science / Virology / viruses / Writing / Zoology