Development and Validation of a Pan-Genotypic Real-Time Quantitative Reverse Transcription-PCR Assay To Detect Canine Distemper Virus and Phocine Distemper Virus in Domestic Animals and Wildlife
Geiselhardt F. Peters M. Jo W.K. Schadenhofer A. Puff C. Baumgärtner W. Kydyrmanov A. Kuiken T. Piewbang C. Techangamsuwan S. Osterhaus A.D.M.E. Beineke A. Ludlow M.
May 2022American Society for Microbiology
Journal of Clinical Microbiology
2022#60Issue 5
Canine distemper virus (CDV) is an animal morbillivirus belonging to the family Paramyxoviridae and has caused major epizootics with high mortality levels in susceptible wildlife species. In recent years, the documented genetic diversity of CDV has expanded, with new genotypes identified in India, the Caspian Sea, and North America. However, no quantitative real-time PCR (RT-qPCR) that has been validated for the detection of all genotypes of CDV is currently available. We have therefore established and characterized a pan-genotypic probe-based RT-qPCR assay based on the detection of a conserved region of the phosphoprotein (P) gene of CDV. This assay has been validated using virus strains representative of six genotypes of CDV in different sample types, including frozen tissue, formalin-fixed paraffin-embedded tissue sections, and virus isolates. The primers and probe target sequences were sufficiently conserved to also enable detection of the phocine distemper virus strains responsible for epizootics in harbor seals in the North Sea in 1988 and 2002. Comparison with two recently published RT-qPCR assays for CDV showed that under equivalent conditions the primers and probe set reported in this study were more sensitive in detecting nucleic acids from an Asia-4 genotype, which displays sequence variation in primer and probe binding sites. In summary, this validated new pan-genotypic RT-qPCR assay will facilitate screening of suspected distemper cases caused by novel genotypes for which full genome sequences are unavailable and have utility in detecting multiple CDV strains in geographical regions where multiple genotypes cocirculate in wildlife. Copyright
canine distemper virus , domestic animals , genotypes , Morbillivirus , pan-genotypic , phocine distemper virus , RT-qPCR , wildlife
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Research Center for Emerging Infections and Zoonoses, University of Veterinary Medicine (TiHo-RIZ), Hannover, Germany
Chemisches und Veterinäruntersuchungsamt Westfalen, Arnsberg, Germany
Department of Pathology, University of Veterinary Medicine, Hannover, Germany
Laboratory of Viral Ecology, Research and Production Center for Microbiology and Virology, Almaty, Kazakhstan
Department of Viroscience, Erasmus Medical Center, Rotterdam, Netherlands
Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand
Animal Virome and Diagnostic Development Research Group, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand
Research Center for Emerging Infections and Zoonoses
Chemisches und Veterinäruntersuchungsamt Westfalen
Department of Pathology
Laboratory of Viral Ecology
Department of Viroscience
Department of Pathology
Animal Virome and Diagnostic Development Research Group
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