猪伪狂犬病毒野毒株与疫苗株的多重PCR鉴别

Multiplex PCR identification of pseudorabies vaccine strains and wild viruses

为建立一种快速鉴别猪伪狂犬病毒野毒株与疫苗株的方法,根据猪伪狂犬病毒(PRV)全基因序列,并参照SA215、Bartha-K61疫苗株基因缺失位点特征,在单一检测野毒株与疫苗株的基础上,针对g E、g B和TK基因设计引物,首次建立了同时检测猪伪狂犬病毒野毒株与疫苗株的多重PCR检测方法。通过一次PCR检测能区分感染野毒株、疫苗株SA215和Bartha-K61,且与猪圆环Ⅱ型病毒(PCV-2)、细小病毒(PPV)、猪繁殖与呼吸综合征病毒(PRRSV)和猪瘟病毒(CSFV)无交叉反应。多次试验证明该方法简单、可靠且敏感度高,可用于科学研究和临床诊断等方面。

In order to establish a diagnostic method for rapid identification of swine pseudorabies virus(PRV)wild virus infection and vaccine strains,this study was developed based on the whole gene sequence of PRV and the characteristics of gene deletion sites of SA215 and Bartha-K61 vaccine strains.On the basis of single detection of vaccine virus and wild virus,primers designed for the three genes of gE,gB and TK were used to establish a multiplex PCR assay for the simultaneous detection of wild virus infections and vaccine strains in pigs.The infection of wild-type virus,vaccine strains SA215 and Bartha-K61 were distinguished by the PCR assay once and there was no cross reaction with porcine circovirus type 2(PCV-2),porcine parvovirus(PPV),porcine reproductive and respiratory syndrome virus(PRRSV)and classical swine fever virus(CSFV).Several experiments have proved that the method is simple,reliable and highly sensitive.It can be used in scientific research,clinical diagnosis and other aspects.