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作者在用紫外线(UV)照射增殖于人胚胎成纤维细胞(HFS)的纯化的甲型肝炎病毒(HAV)后,对其抗原性、感染性、基因组滴度作了评估,并与现行的福马林(FM)灭活法进行了比较。 毒株HAV/HFS/GBM在HFS传30代后,用胰蛋白酶消化细胞单层。离心收集细胞,用1％NP40复悬,反复冻融3次。再于冰浴中声波处理5分钟,离心除去细胞碎片,用聚乙二醇浓缩上清,对生理盐水透析,抽提两次,CsCl梯度离心22小时,从1.23～1.33g/cm~3处收集HAV,对生理盐水透析,用0.2μm滤膜过滤。纯化的HAV用0.9％NaCl溶液稀释100倍。用最大强度接近254nm的低压
The authors evaluated their antigenicity, infectivity, and genomic titers following UVA irradiation of purified Hepatitis A virus (HAV) that proliferated in human embryonic fibroblasts (HFS) and compared them with current blessing Marin (FM) inactivation method was compared. Strains HAV / HFS / GBM After 30 passages of HFS, cell monolayers were trypsinized. Cells were collected by centrifugation, resuspended with 1% NP40, and thawed repeatedly 3 times. Sonicated for 5 minutes in an ice bath, and centrifuged to remove cell debris. The supernatant was concentrated with polyethylene glycol, dialyzed against physiological saline, extracted twice, and centrifuged for 22 hours in a CsCl gradient from 1.23 to 1.33 g / cm 3 HAVs were collected, dialyzed against saline and filtered through a 0.2 μm filter. Purified HAV was diluted 100-fold with 0.9% NaCl solution. With the maximum intensity close to 254nm low pressure