Differentiation of canine distemper virus isolates in fur animals from various vaccine strains by reverse transcription-polymerase chain reaction-restriction fragment length polymorphism according to phylogenetic relations in china

Abstract

In order to effectively identify the vaccine and field strains of Canine distemper virus (CDV), a new differential diagnostic test has been developed based on reverse transcription-polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP). We selected an 829 bp fragment of the nucleoprotein (N) gene of CDV. By RFLP analysis using BamHI, field isolates were distinguishable from the vaccine strains. Two fragments were obtained from the vaccine strains by RT-PCR-RFLP analysis while three were observed in the field strains. An 829 nucleotide region of the CDV N gene was analyzed in 19 CDV field strains isolated from minks, raccoon dogs and foxes in China between 2005 and 2007. The results suggest this method is precise, accurate and efficient. It was also determined that three different genotypes exist in CDV field strains in fur animal herds of the north of China, most of which belong to Asian type. Mutated field strains, JSY06-R1, JSY06-R2 and JDH07-F1 also exist in Northern China, but are most closely related to the standard virulent strain A75/17, designated in Arctic and America-2 genetype in the present study, respectively.

Phylogenetic analysis of the haemagglutinin gene of canine distemper virus strains detected from giant panda and raccoon dogs in China

“…The H gene-based neighbor-joining phylogenetic tree analysis using selected sequences retrieved from GenBank revealed geographic-related patterns of segregation. Our findings are consistent with previous studies demonstrating the occurrence of strains belonging to genetically distinct CDV lineages in Asia [20,28,29]. The Chinese Asia-1 strains displayed a high genetic conservation with other Asia-1 CDVs, which detected from raccoon dogs or other carnivores in China over nearly two decades (Figure 1).…”

Detection and differentiation of field and vaccine strains of canine distemper virus using reverse transcription followed by nested real time PCR (RT-nqPCR) and RFLP analysis

“…Furthermore, the nucleotide sequence differences could be used in RFLP analysis to differentiate vaccine strains from field samples. A similar procedure, using RFLP analysis of the NC gene, was proposed previously to differentiate between vaccine and field strains in China (Wang et al, 2011). Other studies have proposed RFLP analysis for CDV differentiation, although all of them involved the more variable haemagglutinin (H) gene as the target (Mochizuki et al, 1999;Calderon et al, 2007;Di Francesco et al, 2012).…”

Real-time reverse transcription polymerase chain reaction method for detection of Canine distemper virus modified live vaccine shedding for differentiation from infection with wild-type strains

“…3 Many of these differentiating assays are RT-PCR reactions, combined with restriction fragment length polymorphism. 2,4,6,22 While effective, this is time consuming and gelbased PCR assays are not as sensitive as real-time RT-PCR assays. Gel-based nested PCR tests can be as sensitive as real-time PCR tests, and some of the published protocols are nested reactions.…”

“…This quantitative information allows discrimination of vaccine interference from infection with a wild-type strain of distemper virus as the amount of virus present during infection is typically exponentially more than would be detected because of recent vaccination There have been multiple gel-based PCR protocols designed to distinguish vaccine strains from wild-type strains. [2][3][4]6,15,20,22,24 Such tests are designed to take advantage of sequence differences between the vaccine strains and wildtype strains. Most of the tests amplify regions of the hemagglutinin (H) gene 4,6,15,24 or the matrix gene-fusion gene (M-F) intergenic region, 3,20 which have been discovered to be variable among CDV strains.…”