一株猪繁殖与呼吸综合征病毒的分离及ORF5基因序列与国内外毒株的分析和比较  

Isolation of Porcine Reproductive and Respiratory Syndrome Virus Field Strain and Sequence Analysis of ORF5 Gene in Comparison with Other Isolates

作　　者：李晓静[1] 周恩民[1] 赵钦[1] 肖一红[1] 马利宏[1] 

机构地区：[1]山东农业大学动物医学院免疫生物学研究室,山东泰安271018

出　　处：《西南农业学报》2009年第4期1103-1109,共7页Southwest China Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(30871857);山东省自主创新成果转化重大专项项目(2008ZHZX1A1101);山东农业大学"泰山学者"基金项目

摘　　要：分析一株猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)ORF5基因遗传变异情况及与国内外其他毒株比较为指导当地生产提供一定依据,通过常规病毒检测和分离方法,从2008年山东无名高热病猪送检样品肺部组织中检测并分离PRRSV。根据PRRSV美洲经典株VR-2332的ORF5基因设计特异引物,利用RT-PCR和基因克隆ORF5基因,分析其与国内外PRRSV毒株差异。经过细胞病变和间接免疫荧光检测证明所分离到的PRRSV是一山东地方流行毒株,命名为SD-4。通过分析ORF5基因序列及与其它毒株的比较,将北美洲株分为Ⅰ亚群和Ⅱ亚群。SD-4的核苷酸与Ⅰ亚群和Ⅱ亚群及欧洲毒株相比同源性分别为87.4%～89.9%,92.7%～99.2%和63.7%～63.8%。其氨基酸序列的同源性相比分别为85.1%～91.0%,90.5%～99.5%和56.7%～57.7%。进化树分析结果表明SD-4与高致病性毒株JXA1,HUB1,HEB1和SY0608的亲缘关系很近。PRRSV山东分离株SD-4属于美洲经典毒株,可能为高致病性PRRSV毒株。To analyze the ORF5 mutation of one porcine reproductive and respiratory syndrome virus （PRRSV） isolated in Shandong province and to compare this isolated strain with other PRRSV strains, standard virological methods were employed to detect and isolate PRRSV from lung tissue samples collected from pigs with high fever and severe respiratory syndrome in 2008. The primers for ORF5 gene of PRRSV North American （NA） strain VR-2332 were synthesized to clone ORF5 gene by RT-PCR. The cloned ORF5 gene was sequenced and analyzed in comparison with PRRSV isolates reported in GenBank. Based on the cytopathic effect and IFA test,the PRRSV was a filed isolate in Shan- dong province and designated SD-4. From the ORF5 gene sequence analysis results ,PRRSV NA strains can be divided into two subgroups. SD-4ORF5 gene shares 87.4 % - 89.9 % ,92.7 % - 99.2 % and 63.7 % - 63.8 % uucleotide identity with subgroup Ⅰ,Ⅱand European strain,respectively; and the amino acid sequence homology are 85.1% - 91.0 % ,90.5 % - 99.5 % and 56.7 % - 57.7 %. Phylogenetic analysis based on the ORF5 gene showed that SD-4 was more close to JXA1 ,HUB1 ,HEB1 and SY0608 ,which were identified as highly pathogenic PRRSVs. SD-4 isolate belongs to PRRSV NA strain subgroup Ⅱ and was a possible highly pathogenic PRRSV strain.

关 键 词：猪繁殖与呼吸综合征病毒 PRRSV ORF5 同源性 

分 类 号：S858.28[农业科学—临床兽医学]