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钱汶, 陈军, 金巧军, 张轶侠, 陈悦青, 夏华良, 庄成, 邵健忠. 冻干甲型肝炎减毒活疫苗成品检定方法的优化[J]. 中国公共卫生, 2006, 22(11): 1359-1361.
引用本文: 钱汶, 陈军, 金巧军, 张轶侠, 陈悦青, 夏华良, 庄成, 邵健忠. 冻干甲型肝炎减毒活疫苗成品检定方法的优化[J]. 中国公共卫生, 2006, 22(11): 1359-1361.
QIAN Wen, CHEN Jun, JIN Qiaojun, . Optimization of testing method for final product of freezed-dried live attenuated hepatitis A vaccine[J]. Chinese Journal of Public Health, 2006, 22(11): 1359-1361.
Citation: QIAN Wen, CHEN Jun, JIN Qiaojun, . Optimization of testing method for final product of freezed-dried live attenuated hepatitis A vaccine[J]. Chinese Journal of Public Health, 2006, 22(11): 1359-1361.

冻干甲型肝炎减毒活疫苗成品检定方法的优化

Optimization of testing method for final product of freezed-dried live attenuated hepatitis A vaccine

  • 摘要:
      目的  对冻干甲肝减毒活疫苗成品检定中关键性项目的检定方法进行优化。
      方法  (1) 感染性滴度检测:建立微量免疫荧光法, 并与常规组织培养ELISA法比较; (2)牛血清白蛋白残留量检测:比较反向间接血凝法、酶联免疫法的重复性、精确性; (3)水分含量测定:建立助溶剂加入法, 并比较助溶剂加入前后冻干疫苗水分含量检测结果。
      结果  (1) 微量免疫荧光法灵敏度与常规组织培养ELISA法相仿(P >0.05), 重复性佳, 可提前1周左右出结果; 可用Leica Qwin荧光定量分析系统标定化判断结果。(2)反向间接血凝法测定结果不稳定, 2批疫苗5次测定结果的变异系数(CV)为39.12%, 34.23 %; 酶联免疫法的重复性, 精确性均较佳(CV)为2.12 %, 2.82 %, 回收率为101.73%。(3)助溶剂加入前后2批疫苗5次测定结果的CV分别由8.32 %, 9.03%降为1.85%, 1.82 %。
      结论  微量免疫荧光法有望代替常规组织培养ELI SA法用于疫苗滴度检测; 牛血清白蛋白残留量测定中酶联免疫法优于反向间接血凝法; 加入助溶剂有利于提高疫苗水分含量检测的重复性。

     

    Abstract:
      Objective  To optimize some key items in testing method for final product of freezed-dried live attenuated hepatitis Avaccine.
      Methods  Testing method for in fective titer:To establish micro-immunofluorescent method and compare its advantages with that of conventional cell culture ELISA method; Testing method for bovine serum protein residual content:To compare the reproducibility and accuracy between reverse indirecthemagglutination method and ELISA method; Testing method for water content:To establish "solvent-aid adding" method and the vaccine water content results were compared before and after solvent-aid adding.
      Results  (1) The sensitivity of the micro-immunofluorescent method and conventional cell culture ELISA method was in same level(P>0.05), but the former reproducibility was better.The results could be judged one week earlier; Leica Qwin fluorescence analysis software was helpful in getting standardization results.(2) The results of reverse indirecthemagglutination method were not stable, the variation coecficient value(CV)were 39.12% and 34.23% in two batches vaccine respectively(5 times testing).The accuracy and reproducibility of ELISA method we, reall better(CV 2.19%, 2.82%, reclamation 101.73%).(3)The CVin two batches vaccine results were decreased from 8132%, 9.03% to 1.85%, 1.82% respectively before and after the solvent-aid adding(5 times testing).
      Conclusion  Micro-immunofluorescent method may replace the routine cell culture ELISA method in infective titer testing; ELISA method is better than reverse indirecthemagglutination method for bovine serum protein residual content testing; The adding of solvent-aid can improve the reproducibility of water content testing.

     

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