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杨净思, 胡云章, 蒋国润, 杨晓蕾, 柯华昕, 杨伟, 杨蓉. 甲型肝炎减毒活疫苗生产中病毒释放的最佳方法探索[J]. 中国公共卫生, 2001, 17(8): 687-688. DOI: 10.11847/zgggws2001-17-08-10
引用本文: 杨净思, 胡云章, 蒋国润, 杨晓蕾, 柯华昕, 杨伟, 杨蓉. 甲型肝炎减毒活疫苗生产中病毒释放的最佳方法探索[J]. 中国公共卫生, 2001, 17(8): 687-688. DOI: 10.11847/zgggws2001-17-08-10
YANG Jing-si, . Best Method to Release Virus in the Production of Live Attenuated Hepatitis A Vaccine[J]. Chinese Journal of Public Health, 2001, 17(8): 687-688. DOI: 10.11847/zgggws2001-17-08-10
Citation: YANG Jing-si, . Best Method to Release Virus in the Production of Live Attenuated Hepatitis A Vaccine[J]. Chinese Journal of Public Health, 2001, 17(8): 687-688. DOI: 10.11847/zgggws2001-17-08-10

甲型肝炎减毒活疫苗生产中病毒释放的最佳方法探索

Best Method to Release Virus in the Production of Live Attenuated Hepatitis A Vaccine

  • 摘要: 目的探索甲型肝炎减毒活疫苗(简称甲肝活疫苗)生产中感染细胞释放病毒的最佳方法.方法采用超声波细胞粉碎仪、-70℃及液氮冻融3种方法粉碎细胞,对细胞破碎情况、病毒抗原滴度和感染性滴度进行了比较.结果超声波连续粉碎3次(90秒/次)、-70℃反复冻融粉碎4次和液氮冻融粉碎6次,细胞破碎率分别为94%、82%和81%;病毒抗原滴度和感染性滴度均达高峰;抗原滴度均为1:256,感染性滴度(logCCID50/ml)分别为8.67、6.5和6.0.继续超声和-70℃冻融粉碎细胞,病毒抗原滴度和感染性滴度基本不变,而液氮冻融至第10次时,抗原滴度无明显变化,感染性滴度下降2.0logCCID50/ml.结论超声波破碎率最高,细胞碎片最小,病毒充分释放,且不损伤病毒;-70℃和液氮冻融破碎率较低,细胞碎片较大,病毒释放效率低,病毒损伤以液氮冻融为重.

     

    Abstract: ObjectivesIn order to explore the most efficient release of hepatitis A virus(HAV)H2 Strain from the human embryonic lung diploid fibroblast(KMB17)in vaccine production.MethodsThree methods(ultrasonic,deep low temperature freeze-thawing(-70℃)and liquid nitrogen freeze-thawing)were compared according to the index:the rate of celullar lysis,antigenic titer and infectious titer.ResultsBy continuous ultrasonic for three times(90 seconds per time)being handled for four times by freeze-thawing at-70℃,being put into liquid nitrogen for six times,the rate of cellular lysis reached 94%,82%,81% respectively,the antigentiter all were 1:256(log CCID50/ml),infectious titer were 8.67,6.5,6.0 respectively.Further freeze-thawing inliquid nitrogen showed no change in antigen titers,but virus infectious titer decreased down to 210 log CCID50/ml,at the 10th time.ConclusionThe results showed that the ultrasonic is best method to release the virus and do minimal damage to viral proteins and RNA among three ones.

     

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