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病死猪圆环病毒2型检测及全基因序列分析

点击数:2787  时间:2019-10-24 15:23:37  来源: 信息处      作者: 高志强,汪 琳,赵相鹏 等

为准确检测猪圆环病毒2型(PCV2)并分析其基因序列情况,在序列比对分析的基础上,设计一对简并引物和一条探针,经体系优化建立了PCV2的荧光PCR方法。该方法可检出10拷贝的PCV2质粒DNA,不与猪瘟病毒等多种病毒发生交叉反应。对送检的2批次45份病死猪脏器进行检测,检出25份阳性样品,与套式PCR方法的复合率为95.6%;对3份强阳性样品使用一对全基因组扩增引物,扩增了PCV2型全基因片段;经克隆测序后分析,发现3个病毒基因分别属于2种基因亚型(PCV2bPCV2d)。本研究对于PCV2的准确检测及其变异特征的了解具有参考意义。

Detection ofPorcine Circovirus Type 2 in Dead Pigs and Analysis on ItsComplete Genome Sequence

In order toaccurately detect porcine circovirus type 2PCV2and to analyze its genome sequencea pair of degenerate primers and aprobe were designed based on sequence alignment analysisafter systematic optimizationa real-time PCR method was establishedwhich could detect 10 copies ofplasmid DNA of PCV2with no anycross reaction with classical swine fever virus and other viruses. Then twobatches of 45 organ samples collected from dead pigs were tested by establishedmethodand 25 samples were detected to bepositivethe coincidence rate with nested-PCRwas 95.6%for the three strong-positive samplestheir complete gene fragments of PCV2were amplified by a pair of primersthen weresequenced and analyzedand it wasfound that the genes of the three viruses were classified into two differentsubtypesPCV2b and PCV2d. Thereforeit was of certain significance toconfirm relevant test results and recognize any variation of PCV2 by the abovemethod.

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