鉴别PCV2b与PCV2d基因亚型PCR方法的建立及应用

Chinese Veterinary Science(2017)

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Abstract
为建立可鉴定PCV2b和PCV2d基因亚型的PCR方法,通过分析GenBank内PCV2ORF2基因序列,设计了可分别扩增ORF2基因和鉴别PCV2b、PCV2d的引物,并建立PCR方法.应用PCV2b和PCV2d引物扩增108份PCV2阳性样本,选择11份PCV2b和19份PCV2d单一阳性样本扩增ORF2基因序列,并用于遗传进化分析.结果显示,PCV2b和PCV2d引物扩增的敏感性较好,PCV2b和PCV2d最低可扩增基因拷贝数分别为19 copies/μL和80 copies/μL.108份PCV2阳性DNA中PCV2b阳性率为42.5%,PCV2d阳性率为69.4%,混合感染阳性率为12%.30条ORF2序列的遗传进化分析结果显示,11条为PCV2b基因亚型,19条为PCV2d基因亚型,遗传进化分析结果与PCR结果一致.本试验建立了一种鉴别PCV2b和PCV2d的PCR方法,可用于PCV2流行病学的调查.
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