PRRSV强弱病毒株嵌合感染性克隆的构建及鉴定  

The construction and rescue of chimeric highly pathogenic PRRSV and attenuated PRRSV

作　　者：吕健[1,2] 韦祖樟[1] 高飞[1] 郑海红[1] 童光志[1] 袁世山[1] 

机构地区：[1]中国农业科学院上海兽医研究所,上海200241 [2]国家知识产权局专利审查协作北京中心,北京100193

出　　处：《中国预防兽医学报》2013年第4期263-266,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30972204);青年科学基金(30901078)

摘　　要：为构建高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)与致弱PRRSV的重组病毒,本研究在PRRSV致弱株APRRS的全长感染性克隆pAPRRS与HP-PRRSV感染性克隆pJX143的基础上,利用SOE-PCR技术将强弱毒的nsp2区域进行互换,构建了nsp2基因替换的强弱毒PRRSV嵌合感染性克隆。将构建的嵌合克隆转染MA104细胞,4 d后观察到典型的细胞病变。通过RT-PCR和间接免疫荧光证明获得了强弱病毒株之间nsp2基因互换的嵌合病毒。这些嵌合病毒的构建和相应反向遗传操作平台的建立,为进一步研究HP-PRRSV的致病机制及相关疫苗的研发奠定了基础。In order to construct recombinant virus between the highly pathogenic porcine reproductive and respiratory syndrome virus （HP-PRRSV） and the attenuated virus strain, based on an infectious cDNA clone of an attenuated PRRSV strain pAPRRS and the HP-PRRSV eDNA clone pJX143, we replaced the coding sequence of pAPRRS nsp2 with those of the HP-PRRSV by SOE-PCR. Upon transfection of recombinant virus into MA104 cells, typical PRRSV cytopathic effects were observed at 4 day post transfection. The rescued chimeric PRRSVs were identified by RT-PCR and indirect immunofluorescence assay. This study provided a valuable tool to develop the chimeric PRRSV as vaccine candidate offering cross-protection to HP-PRRSVs and to investigate the mechanism of pathogenesis of the increasing-virulence of the on-going prevalent PRRSV in China.

关 键 词：PRRSV 感染性克隆 嵌合病毒 病毒株 猪繁殖与呼吸综合征病毒 PCR技术 鉴定 间接免疫荧光 

分 类 号：S858.285.3[农业科学—临床兽医学]