TGEV细胞培养物超速离心与RT-PCR扩增的研究  

Ultra Centrifugation the Cell Cultures of TGEV and RT-PCP

作　　者：蒋凤英[1] 何锡忠[1] 王英[1] 李春华[1] 钱永清[1] 邹勇[1] 

机构地区：[1]上海农科院畜牧兽医研究所,上海201106

出　　处：《动物医学进展》2007年第4期50-52,共3页Progress In Veterinary Medicine

基　　金：上海农业"四新"技术推广项目[沪农科推字(2005)第2-2号]

摘　　要：研究猪传染性胃肠炎病毒(TGEV)细胞培养物的超速离心及鉴定方法。将TGEV的细胞培养物用饱和硫酸氨法浓缩后,用200 g/kg^500 g/kg蔗糖密度梯度离心分装,测定蛋白浓度并取峰值管进行RT-PCR,并设立15、25、35三个不同循环参数测定PCR产物volume值。结果表明,在35个循环条件下第14号样品PCR产物的volume值最大,表明该样品病毒核酸的含量最大,由此推算病毒粒子主要集中在360 g/kg^380 g/kg蔗糖区带处,这与理论值相符,证明猪传染性胃肠炎病毒经密度区带离心后,利用PCR产物量确定目的病毒区带效果显著。The article focused on the ultra centrifugation and identification method of TGEV cells. TGEV cells were concentrated by saturation ammonium sulfate sedimentation, and then isolated by discontinuous sucrose density gradient （200 g/kg-500 g/kg）. The samples were separated automatically in different tubes,and concentration of protein were detected. The tube with peak value was used for RT-PCR. Three different cycle parameters （15,25 and 35） were used for detecting volume of outcome of RT-PCR. The result showed that in the condition of 35 cycles, the highest volume is No. 14 tube. It is concluded that the coronavirus is enriched in 360 g/kg-380 g/kg sucrose density, and it is identical with theory. It proved that it got marked effect by centrifugation of TGEV and RT-PCR.

关 键 词：猪传染性胃肠炎病毒 蔗糖密度梯度离心 RT-PCR 

分 类 号：Q78[生物学—分子生物学]