摘要
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。
Eight cDNA fragments overlapped each other have be en obtained from th e live attenuated hepatitis A vaccine K 7,the progeny viruses of H 2 strain b y R T-PCR.A total of 21 fragments were sequenced with the dye termiantor cycle seque ncing kit and automatic sequence equipment model 377(ABI corp.).By using the sof tware of Maxtrigene,the complete nucleotide sequence of K 7 cDNA has been deter mined and analyzed.An entire length of 7443 bases was evidenced for the s equence of K 7 virus with homology rate of 99.75%,99.61% and 99.95% against its parenta l H 2 atrain,HM 175 strain and the attenuated HM 175 (HM 175 ATD) .According to the d ifferences in the 168 bases located at the joint site of P1 and P2 region,the ge notype of K 7,H 2 as well as HM 175 should be classified into ⅠB subgeno type.The K 7,H 2 and HM 175 ATD have showed the same deletion of nucleotide 131 1 34 and 203 at the 5'non encoding area,and also the identical sequence for encoding the s tructural protein.The result of this study has demonstrated the high purity of K 7 vaccine with the technique of molecular virology and provided the basic info rm ation for carrying out the molecular epidemiologic investigation for hepatitis A .
出处
《云南大学学报(自然科学版)》
CAS
CSCD
1999年第3期170-174,共5页
Journal of Yunnan University(Natural Sciences Edition)
基金
美国中华医学基金
关键词
甲肝病毒
甲肝减毒活疫苗
核苷酸序列
测定
hepatitis A virus
live attenuated hepatiti s A vaccine
nucleotide sequence
genotype