TGEV细胞培养物超速离心与RT-PCR扩增的研究

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.

Ultra Centrifugation the Cell Cultures of TGEV and RT-PCP

Abstrat: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.