猪伪狂犬病毒Real-time PCR检测方法的建立和弱毒疫苗病毒含量的检测

Establishment of Real-time PCR Assay of Pseudorabies Virus and Viral Load Detection of PRV Attenuated Vaccine

根据伪狂犬病毒的gB基因设计引物,将含gB基因的质粒作为标准品,绘制标准曲线,建立了伪狂犬病毒PRV的荧光定量PCR检测方法。该检测方法具有良好的重复性、敏感性和特异性,不与PCV2、PPV、CSFV、JEV和PRRSV发生反应。用该方法对不同滴度病毒液的基因拷贝数进行检测,发现病毒滴度和病毒拷贝数之间存在较好的相关性。采用该方法对7种商品化PRV弱毒疫苗进行病毒含量检测,结果显示不同商品化弱毒疫苗的病毒含量存在显著差异,与说明书标注的病毒含量基本一致。

In order to establish a real-time PCR method for detecting gB gene of PRV,the author designed a pair of primers and analyzed the real-time PCR method by different quantitive concentration of plasmids containing PRV gB gene.A standard curve was achieved.The results showed that this method had good repeatability,specificity and sensitivity,and did not react with PCV2,PPV,CSFV,JEV and PRRSV.The relationship of viral titer and gene copies of different PRV dilutions was analyzed by the method.The viral titer had a better correlation with viral copies.The method was used to detect the viral content of seven kinds of commercial attenuated PRV vaccines.The viral content of different commercial attenuated PRV vaccines was significantly different,which was basically consistent with the instructions.