H2株甲肝减毒活疫苗K7的核苷酸全序列分析

应用ＲＴ－ＰＣＲ获取Ｈ２株甲肝减毒活疫苗（Ｋ７）的ｃＤＮＡ片段，经末端终止测序ＰＣＲ试剂盒和全自动测序仪（ＡＢＩ３７７）作序列测定，用Ｍａｘｔｒｉｇｅｎｅ软件进行计算机分析，Ｋ７疫苗病毒的核苷酸全长含７４４３个碱基．Ｋ７对比其亲代Ｈ２株，减毒的ＨＭ１７５株以及ＨＭ１７５野毒株，同源性分别为９９．７５％，９９．９５％和９９．６１％；彼此间在Ｐ１和Ｐ２连接区１６８个碱基的同源性为９９．４％～１００％，均为ⅠＢ基因亚型．前三者在５’非编码区均有１３１～１３４位和２０３位５个碱基缺失，且结构蛋白基因区序列完全一致．本文用分子病毒学方法证实了Ｋ７疫苗的优异纯毒水平，也为开展甲型肝炎分子流行病学工作提供了基础资料     

STUDIES IN MONKEYS OF ATTENUATED HEPATITIS A VARIANTS

Three HAV variants, H2M20(35℃), H2M20(32℃) and H2M20K5(32℃), were developed by passagein different tissue culture cells and at different temperatures. Virulence for monkeys was assessed by in-oculating each of the variants into four monkeys. Weekly bleeding through 16 weeks post-inoculation wasassayed for anti-HAV titers and SGPT as well as LDH5 activities. And in some monkeys liver biopsieswere also studied. Seroconversion was induced in all of the inoculated monkeys. The anti-HAV titers test-ed at 16 weeks post-inoculation were: 40-640 with H2M20(35℃), 40-160 wi1h H2M20(32℃) and 20-40 with H2M20K5(32℃). These variants showed different levels of virulence/attenuation for monkeys.Variant H2M20(35℃) showed no evidence of attenuation, variant H2M20(32℃) retained slight virulenceand variant H2M20K5(32℃) showed no evidence of virulence for monkeys. Criteria are proposed for as-say of HAV virulence tested in monkeys.     

H2株甲肝减毒活疫苗K7的核苷酸全序列分析

应用RT-PCR获取H     

甲型肝炎减毒活疫苗的研究——Ⅰ.不同毒力毒株的猴体试验

三株减毒水平不同的甲型肝炎病毒株H2M20(35℃),H2M20(32℃),H2M20,K5(32℃),各接种4只恒河猴,3—10周后甲肝抗体全部阳转,接种后16周的抗体滴度:H2M20(35℃)达1:160—1:640,H2M20(32℃)为1:40—1:160,H2M20K5(32℃)为1:20—1:40。猴接种病毒后16周内,每周检测了SGPT,LDH5,部分猴进行了肝穿活组织检查。结果说明,H2M20(35℃)属强毒力反应,而H2M20K5(32℃)接种的4只恒河猴未观察到有任何肝炎指征,属弱毒株反应。在毒力和抗体反应方面,H2M20K5(32℃)是值得进一步研究的甲型肝炎减毒疫苗候选株。文中提出了恒河猴检定甲肝病毒毒力反应的判断标准。