Establishment and evaluation of a multiplex real-time RT-PCR for quantitative and differential detection of wild-type canine distemper virus from vaccine strains

This study sought to establish a real-time reverse transcription (RT)-PCR method differentially detect canine distemper virus (CDV) wild-type and vaccine strains. To this end, pair of CDV universal primers two specific minor groove binder (MGB) probes, harboring T/C substitution in the hemagglutinin (H) gene, were designed. Using recombinant plasmid expressing H gene or strain as standards, sensitive multiplex RT-PCR was established for quantitative differential detection The limit assay 22.5 copies/μL 2.98 viral RNA strains, respectively. Importantly, MGB probes specifically hybridized different genotypes circulating China well globally administered viruses, respectively, with no cross-reactivity observed non-CDV viruses. Moreover, successfully applied tissue samples experimentally infected breeding foxes, raccoon dogs, minks. Additionally, able whole blood early 3 days post-infection, 4 prior onset clinical signs these infection animals. Hence, is useful differentiating strains China, conducting diagnosis dynamic mechanism replication studies vivo.