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

Abstract

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.

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

“…A pan-genotypic Taq-Man-based RT-qPCR assay has been designed and validated for detection of all genotypes of CDV, and the primers and probe bind to a conserved region of the P gene, enabling amplification of a 160bp product from all described CDV and PDV genotypes. However, this assay can not be used for the identification of wild-type and vaccine CDV strains [ 19 ]. Currently, the use of vaccines has helped prevent CD, it has also created difficulties associated with distinguishing between CDV wild-type and vaccine strains in infected animals.…”

“…To validate the agreement of samples that underwent multiplex real-time RT-PCR and other assays, 75 samples suspected CDV positive tissues samples were collected from dogs and fur-bearing animals from Shandong, Liaoning, Hebei, and Jilin provinces in China and subjected to the multiplex real-time RT-PCR [ 19 ] and the “RT-PCR-RFLP assays” as described previously [ 20 ].…”