3株PRRSV变异株理化特性研究及NSP2、ORF5基因序列分析  

Study of physicochemical property of three variant PRRSV strains and sequence analysis of NSP2 and ORF5 genes

作　　者：王连想[1,2] 孙彦伟 马静云[2] 曹永长[3] 任裕其 田云 谢青梅 毕英佐[2] 于康震[4] 

机构地区：[1]广东省动物防疫监督总所,广东广州510230 [2]华南农业大学动物科学学院,广东广州510642 [3]中山大学生命科学学院,广东广州510275 [4]中国兽医药品监察所,北京100081

出　　处：《中国兽医学报》2010年第5期565-568,共4页Chinese Journal of Veterinary Science

基　　金：广东省科技计划基金资助项目(2005A20902001);广东省农业厅2007年动植物防疫检疫研究专项基金资助项目

摘　　要：用Marc-145细胞从广东省3个发病猪群分离到3株病毒,经RT-PCR方法鉴定为PRRSV变异株(NSP21594～1680bp缺失)。3株分离株第1代均对Marc-145细胞适应性不强,第2代可致典型病变,第3代TCID50分别为103.67、104.20、104.75mL-1;在透射电镜下观察主要为40～70nm大小的圆形、椭圆形具有囊膜的病毒粒子;均对氯仿、酸、碱和热敏感。3株分离株NSP2基因与CH-1a、VR-2332等经典PRRSV相比均存在2个位点30个氨基酸缺失,与JXA1、HUN4、HPDEBV等变异株具有相同缺失特性且高度相似,相似性为98.1%～98.3%。ORF5基因均由200个氨基酸组成,与JX-A1、HUN4、HPDEBV等变异株高度相似,相似性为98.5%～99.0%,且第13、151位均为具有强毒特性的精氨酸(R)。本研究进一步证实在广东省内出现以NSP2缺失30个氨基酸为特征PRRSV变异株的流行,也为科学监测和综合防控该毒株感染引起的PRRS奠定了基础。Three viruses were isolated by Marc-145 cell from sick and dead pigs in three different farms of Guangdong province and were identified as variant PRRSV strains （NSP2 1 594-1 680 deletion） by RT-PCR.Those variant PRRSV strains could adapt to Marc-145 cell and cause un-severe cell pathological effects （CPEs） at the 1st generation,the CPEs could be easily observed from the 2nd generation,and the TCID50 was 103.67,104.20,104.75mL-1 respectively at the 3rd generation.The expressed virions displayed various shapes including round and ellipse forms with diameters in the range of 40-70 nm.All three isolates were susceptible to chloroform,acid,alkali and heat.The results showed that NSP2 genes of all three strains were highly homologous and had 30 amino acid deletions compared with that of CH-1a and VR-2332,which had the same deletion compared with that of JXA1,HUN4 and HPDEBV.The ORF5 genes were consist of 200 amino acids and were 98.5% to 99.0% of homology with that of JXA1、HUN4 and HPDEBV,and the site of 13 and 151 were Arginine （R） which were related to virulent strains.

关 键 词：猪繁殖呼吸综合征病毒 NSP2基因 ORF5基因 分离 理化特性 

分 类 号：S852.65[农业科学—基础兽医学]