甲型肝炎病毒Vero细胞适应株的选育研究

The study on adaptation of hepatitis A virus YN5 strain in Vero cell

将甲肝患者粪便中分离的甲型肝炎病毒在Vero细胞中进行适应性培养 ,选育高滴度适应株应用于甲肝灭活疫苗研究。在Vero细胞上连续传代 ,测抗原滴度和感染性滴度 ,满意后按WHO推荐的甲型肝炎灭活疫苗规程进行灭活疫苗试制研究。经Vero细胞 14次适应性传代后 ,病毒抗原滴度可高达 1∶2 5 6 0 ,感染性滴度为 8.2 3LogC CID50 /ml。试制的灭活疫苗HPSEC检测在 2 80nm时仅有一个高峰 ,SDS PAGE电泳 ,在 2 2kD、2 6kD和 33kD处有三条蛋白带 ,和HAVVP3、VP2和VP1的位置相同。ICR小鼠效力试验表明疫苗剂量 16 0 0EU/ml与Merck疫苗5 0U效果相似。通过研究获得了Vero细胞甲肝病毒适应株YN5株 ,初步证明可作为甲型肝炎灭活疫苗的候选毒株。

YN5 strain was isolated from feces of hepatitis A patients and adapted to vero cells.The adapted YN5 strain was identified as HAV with the specific experiment.Antigenic titers of YN5 strain P14 on Vero cells were 1∶2560,infective titres of that was 8.23LogCCID 50 /ml.The experimental vaccine was made according to WHO requirement for inactivated hepatitis A vaccine.Experimental vaccine was only one peak through HPSEC at 280nm.By SDS PAGE,YN5 strain contains three peptide components VP1,VP2,VP3,their molecular weights are about 33,26,22kD respectively.In addition,the efficiency of the experimental vaccine,ICR mice immunized with 1600EU/ml was same as Merck vaccine (50U).The research provide YN5 strain adapted Vero cell and was a well strain for producing inactivated vaccine.