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目的 监测H2株甲肝病毒经人胚肺二倍体细胞KMB17培养的毒力 /减毒水平及核苷酸序列。方法H2株甲肝减毒活疫苗H2M2 0K7(K7)用KMB17细胞增殖 ,分别在 35℃和 37℃连续传代后 ,抽查不同代次病毒的普通狨猴接种反应和核苷酸片段序列。结果 H2 KMB17系统在 35℃培育 16代次过程中 ,病毒的抗原滴度和感染性滴度稳定 ,在 37℃的滴度明显低下 ,经 13代次仍未达亲本水平。K18(35℃ ,11代 )和K15 (37℃ ,8代 )病毒经普通狨猴接种反应证实为减毒性质。核苷酸两片段共 1897个碱基的序列分析显示 ,K18和K15与K7的同源性高达 99 3%～ 10 0 %。结论 K7疫苗病毒在KMB17细胞培养经 35℃和 37℃连续传代 ,无毒力回升和遗传稳定性改变
Objective To monitor the virulence / attenuated level and nucleotide sequence of H2 strain Hepatitis A virus cultured in human fetal lung diploid cell KMB17. Methods H2M2 0K7 (K7), an attenuated live attenuated virus of H2 strain, was propagated in KMB17 cells. After being passaged at 35 ℃ and 37 ℃ respectively, the inoculation reaction and nucleotide sequence of common tamarin in different generations of virus were performed. Results During the 16 generations of H2 KMB17 system incubated at 35 ℃, the titer and infectious titer of the virus were stable. The titers at 37 ℃ were significantly lower than those at 13 generations. K18 (35 ℃, 11th generation) and K15 (37 ℃, 8th generation) virus were proved to be attenuated by the common macaque inoculation reaction. Sequence analysis of a total of 1897 nucleotides showed that the homologies of K18 and K15 with K7 were as high as 99 3% ~ 100%. Conclusions The K7 vaccine virus was passaged continuously at 35 ° C and 37 ° C in KMB17 cells with no toxic recovery and altered genetic stability