甲肝病毒在猴胚肾细胞和Vero细胞上的分离与适应

Isolation and Adaptation of HAV on MEK Cells and Vero Cells

使用恒河猴胚肾 (MEK)细胞从临床标本中分离和适应了甲肝病毒W和X ,通过阻断实验、中和实验、免疫荧光和免疫电镜实验、RT PCR对其进行特异性鉴定 ,证明是甲肝病毒。W株和X株第 7代在MEK细胞上的抗原滴度分别为 1∶5 12、1∶10 2 4,感染滴度 (logTCID50 /mL)分别为 8.17、8.5 0。只有分离株W可以适应于Vero细胞 ,第 6代 2 1d抗原滴度为 1∶2 5 6 ,感染滴度 (logTCID50 /mL)为 8.0 0。

HAV strains W and X were isolated and adapted to Rhesus monkey embryo kidney(MEK) cells from feces of hepatitis A patients,and identified to be HAV with blocking experiment,neutralizing experiment,indirect immunofluorescence (IF),immune electron microscopy(IEM) and RT-PCR.Antigenic titers of strains W and X P7 on MEK cells were 1∶512,1∶1024,respectively and infective titers(logTCID 50 /mL)of that were 8.17 ,8.50 respectively.Only the isolate W was adapted to Vero cells.Antigenic titer of adapted strain W P6 on Vero cells on 21th day was 1∶256 and infective titer(logTCID 50 /mL)was 8.00.