速成单层法病毒空斑技术影响因素的探讨

ON FACTORS AFFECTING QUICK MONOLAYER PLAQUE ASSAY OF VIRUSES

速成单层法病毒空斑技术是一种新的病毒空斑技术。本文以脊髓灰质炎病毒Ⅰ型（PV Ⅰ）Mahoney株为代表,探讨速成单层法病毒空斑技术的实验条件。结果表明,选用HeLa细胞、以0.02％EDTA消化、加含20％小牛血清的营养液制成细胞悬液,用国产16孔组织培养板、每孔加细胞4×105/0.4ml,置CO2培养箱吸附贴壁60～90min,并取国产琼脂糖作覆盖物凝胶是速成单层法PV Ⅰ空斑滴定的较佳条件,这对其它病毒-细胞系统的速成单层法空斑试验也具较大的参考价值。

Quick monolayer plaque assay of viruses(QMPAV)is a new technique of plaque assay of viruses.Its optimal expriment conditions,including type and number of cells,tine of virus adsorption and cell attachment,and type of gels and multi-well tissue culture plates were studied with the model of type I poliovirus(PVI),Mahoney strain,The cells disprsed with 0.02％EDTA,suspended in 20％ calf serum medium and cunured in Co_2 incubator were more liable to attached to the wall of culture plates.The plaque titer of the same PV I stock on HeLa cells was higher than that on ZMRUE-85,Hep-2,FL,MERN and MA104 cells,and was generally raised with the incrcasc of cell number or the prolongation of the time of virus adsorption and cell attachment.Four hundred thousand cells per well and attachment for 60 tO 90 minutes are thought to be suitabIe,convenient and economical.The homemade agarose and 16-well culture plates were at last as good as the imported agar powder and 24-well culture plates.thes results were valuable to QMPAV of other virus-cell systems.