中国兽医科学2017，Vol.47Issue(11)：1378-1384,7.DOI:10.16656/j.issn.1673-4696.2017.11.006
鉴别PCV2b与PCV2d基因亚型PCR方法的建立及应用
Development and application of PCR assay for differential detection of porcine circovirus type 2b from 2d
摘要
Abstract
Primers used to amplify the PCV2-0RF2 gene and differentiate PCV2b and PCV2d geno-subtype were designed based on analysis of 0RF2 downloaded from Genbank.The sensitivity and specificity were evaluated using pMD19-T plasmids with PCV2b and PCV2d complete genomes.One hundred and eight PCV2 positive DNAsamples were amplified using PCV2b and PCV2d primers.Complete 0RF2 genome of ii PCV2b and 19 PCV2d single positive DNAwere amplified and sequenced,and phylogenetic trees were reconstructed based on 30 obtained and 34 reference sequences.The specificity of PCV2b and PCV2d primer was well,and the minimum copies of PCV2b and PCV2d were 19 and 80 copies/μ L,respectively.The positive rate of PCV2b and PCV2d in 108 PCV2 DNAsamples was 42.5％and 69.4％,respectively,and the coinfection rate of PCV2b and PCV2d was 12％.Phylogenetic analysis indicated that 110RF2 sequences belonged to PCV2b,while 19 belonged to PCV2d,which was in agreement with PCR results.Conclusion:we designed two primers which could be used for specific amplification of PCV2b and PCV2d.关键词
PCV2/分型检测/分子流行病学调查/遗传分析Key words
PCV2/genotype differentiate/molecular epidemiology investigation/phylogenetic analysis分类
农业科技引用本文复制引用
屈泰龙,严美君,葛猛,李润成,张家祥,余兴龙..鉴别PCV2b与PCV2d基因亚型PCR方法的建立及应用[J].中国兽医科学,2017,47(11):1378-1384,7.基金项目
国家重点研发项目(2017YFD0500102) （2017YFD0500102）
