title: Development of a combined canine distemper virus specific RT-PCR protocol for the differentiation of infected and vaccinated animals (DIVA) and genetic characterization of the hemagglutinin gene of seven Chinese strains demonstrated in dogs.
abstract: A combined reverse-transcription polymerase chain reaction (RT-PCR) method was developed for the detection and differentiation of wild-type and vaccine strains of the canine distemper virus (CDV). A pair of primers (P1/P2) was used to detect both CDV wild-type strains and vaccines. Another pair (P3/P4) was used to detect only CDV wild-type strains. A 335bp fragment was amplified from the genomic RNA of the vaccine and wild-type strains. A 555bp fragment was amplified specifically from the genomic RNA of the wild-type strains. No amplification was achieved for the uninfected cells, cells infected with canine parvovirus, canine coronavirus, or canine adenovirus. The combined RT-PCR method detected effectively and differentiated the CDV wild-type and vaccine strains by two separate RT-PCRs. The method can be used for clinical detection and epidemiological surveillance. The phylogenetic analysis of the hemagglutinin gene of the local wild-type CDV strains revealed that the seven local isolates all belonged to the Asia-1 lineage, and were clustered closely with one another at the same location. These results suggested that the CDV genotype Asia-1 is circulating currently in domestic dogs in China.
authors: Division of Zoonoses, State Key Laboratory of Special Economic Animal Molecular Biology, Institute of Special Wild Economic Animal and Plant Science, Chinese Academy of Agricultural Sciences, 15 Luming Street, Jilin 132109, China. jilinseq@163.com
authors: Li
authors: L
authors: Yi
authors: Shipeng
authors: S
authors: Cheng
authors: Hongli
authors: H
authors: Xu
authors: Jianke
authors: J
authors: Wang
authors: Yuening
authors: Y
authors: Cheng
authors: Shen
authors: S
authors: Yang
authors: Bin
authors: B
authors: Luo
mesh_terms: D000818:Animals; D002681:China; D016000:Cluster Analysis; D017931:DNA Primers / Q000235:genetics; D004216:Distemper / Q000175:diagnosis* / Q000821:virology; D004217:Distemper Virus, Canine / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D004283:Dog Diseases / Q000175:diagnosis* / Q000821:virology; D004285:Dogs; D006388:Hemagglutinins / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000235:genetics; D014765:Viral Vaccines / Q000235:genetics; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers; D006388:Hemagglutinins; D012367:RNA, Viral; D014764:Viral Proteins; D014765:Viral Vaccines