tailieunhanh - Báo cáo y học: " A multiplex reverse transcription-nested polymerase chain reaction for detection and differentiation of wild-type and vaccine strains of canine distemper virus"

A multiplex reverse transcription-nested polymerase chain reaction for detection and differentiation of wild-type and vaccine strains of canine distemper virus | Si et al. Virology Journal 2010 7 86 http content 7 1 86 VIROLOGY JOURNAL SHORT REPORT Open Access A multiplex reverse transcription-nested polymerase chain reaction for detection and differentiation of wild-type and vaccine strains of canine distemper virus Wei Si 1 Shun Zhou 2 Zhao Wang2 and Shang-jin Cui 1 Abstract A multiplex reverse transcription-nested polymerase chain reaction RT-nPCR method was developed for the detection and differentiation of wild-type and vaccine strains of canine distemper virus CDV . A pair of primers P1 and P4 specific for CDV corresponding to the highly conserved region of the CDV genome were used as a common primer pair in the first-round PCR of the nested PCR. Primers P2 specific for CDV wild-type strains were used as the forward primer together with the common reverse primer P4 in the second round of nested PCR. Primers P3 P5 specific for CDV wild-type strain or vaccine strain were used as the forward primer together with the common reverse primer P4 P6 in the second round of nested PCR. A fragment of 177 bp was amplified from vaccine strain genomic RNA and a fragment of 247 bp from wild-type strain genomic RNA in the RT-nPCR and two fragments of 247 bp and 177 bp were amplified from the mixed samples of vaccine and wild-type strains. No amplification was achieved for uninfected cells or cells infected with Newcastle disease virus NDV canine parvovirus CPV canine coronavirus CCV rabies virus RV or canine adenovirus CAV . The RT-nPCR method was used to detect 30 field samples suspected of canine distemper from Heilongjiang and Jilin Provinces and 51 samples in Shandong province. As a result of 30 samples were found to be wild-type-like and 5 to be vaccine-strain-like. The RT-nPCR method can be used to effectively detect and differentiate wild-type CDV-infected dogs from dogs vaccinated with CDV vaccine and thus can be used in clinical detection and epidemiological surveillance. Introduction Canine distemper CD is