PRRSV广东株XH-GD的分离及其NSP2、ORF3、ORF5的序列分析  

Isolation and sequence analysis of NSP2,ORF3 and ORF5 of PRRSV XH-GD strain from Guangdong province

作　　者：曹宗喜[1,2] 潘全会[2] 林哲敏 孔留五[2] 何逸民[2] 张桂红[2] 

机构地区：[1]海南省农科院畜牧兽医研究所/海南省热带动物繁育与疫病研究重点实验室,海南海口571100 [2]华南农业大学农业部兽用疫苗创制重点实验室,广东广州510642

出　　处：《广东农业科学》2013年第12期159-163,共5页Guangdong Agricultural Sciences

基　　金：国家自然科学基金(31272564);国家自然科学基金会(NSFC)-广东联合基金项目(U0931003);国家现代农业产业技术体系专项(CARS-36);海南省科学事业费项目(琼财预[2013]131号)

摘　　要：将采集的广东某猪场疑似猪高热症病料处理后接种于Marc-145细胞,出现细胞聚集、固缩、脱落等特异性细胞病变。根据GenBank中猪繁殖与呼吸综合征病毒(PRRSV)的基因序列设计并合成针对NSP2、ORF3和ORF5基因的引物,扩增目的基因并测序分析。结果表明:分离毒株XH-GD株PRRRSV属于美洲型,且NSP2基因发生30个氨基酸的不连续缺失;XH-GD的NSP2、ORF3和ORF5基因与HUB1、NX06、BJsy06、VR-2332、HB-1(sh)2002、HB-2(sh)2002、CH-la和RespPRRS MLV等毒株等位基因的核苷酸同源性分别为83.3%-98.9%、88.6%-99.2%、88.1%-99.2%,推导的氨基酸同源性分别为76.8%-98.3%、83.7%-98.8%、88.1%-99.2%;而与LV的等位基因核苷酸同源性分别为52.9%、65.0%和64.3%,推导的氨基酸同源性分别为31.3%、57.5%和58.9%。基因系统发育树分析表明,XH-GD与NX06、BJsy06株的亲缘关系很近,与HUB1、NX06、BJsy06、VR-2332、HB-1(sh)2002、HB-2(sh)2002、CH-la和RespPRRS MLV亲缘关系较近,与LV毒株处于不同的分支。The samples collected from Guangdong province were inoculated to Marc-145 cell, the PRRSV specific cytopathogenic effect （CPE） was observed and the isolates named XH-GD obtained. After reverse transcription polymerase chain reaction, the amplified NSP2,0RF3 and ORF5 fragments were analyzed by sequence comparison. The results demonstrated that XH-GD belonged to the North American genotype and its NSP2 had 30 amino acids deletion. The NSP2, ORF3 and ORF5 of XH-GD shared 83.3%-98.9%, 88.6%~ 99.2% and 88.1%-99.2% of nucleotide similarity, and 76.8%-98.3%, 83.7%-98.8% and 88.1%-99.2% of deduced amino acid similarity, with that of HUB1, NX06, BJsy06, VR-2332, HB-1 （sh）2002, HB-2 （sh）2002, CH-la and RespPRRS MLV respectively; and shared only 52.9%, 65.0% and 64.3% of nucleotide similarity, and 31.3%, 57.5% and 58.9% of deduced amino acid similarity with that of LV respectively. The phylogenetic tree revealed XH-GD had closer relationship with HUB1, NX06, BJsy06, VR-2332, HB-1（sh）2002, HB-2 （sh）2002, CH-1a and RespPRRS MLV strains than that of LV strain.

关 键 词：猪繁殖与呼吸综合征病毒 XH—GD毒株 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]