水痘-带状疱疹病毒Oka疫苗株与野毒株序列比对

摘要

目的 观察水痘-带状疱疹病毒(varicella-zoster virus,VZV)Oka疫苗株(V-Oka株)的传代稳定性,并与分离于水痘患者的野毒株进行序列比对.方法 将V-Oka株在人二倍体细胞MRC-5中连续传代培养至50代,提取32、34、38、42、46和50代V-Oka株及野毒株的DNA,对主要基因(ORF31、ORF62、ORF68、R区)进行PCR扩增和测序,并进行序列比对.结果 与GenBank中的V-Oka株序列相比,32代至50代V-Oka株的ORF31、ORF62、ORF68、R区序列稳定,并趋于减毒.未发现野毒株序列发生如同V-Oka株的独特变异.结论 连续传代培养至50代,V-Oka株的分子遗传学特性保持稳定.序列比对显示,水痘患者的致病毒株为野毒株,而非V-Oka株.%Objective To observe the stability of varicella-zoster virus(VZV) Oka vaccine(V-Oka)strain during subculture,and align VZV sequences between V-Oka strain and wild-type strain from chickenpox patients.Methods V-Oka strain was subcultured to the 50th passage on human diploid cell MRC-5.VZV DNAs were extracted from the 32nd,34th,38th,42nd,46th and 50th passages of V-Oka strains and wild-type strains.The major genes( ORF31,ORF62,ORF68 and repeat regions)of V-Oka strains and wild-type strains were amplified by PCR and sequenced.DNA sequences between V-Oka strains and wild-type strains were aligned.Results In comparison with DNA sequences of V-Oka strain in GenBank,ORF31,ORF62,0RF68,repeat regions sequences of V-Oka strains from the 32nd passage to the 50th passage were stable and seemed to be more attenuated.Unique mutations as same as those in V-Oka stains were not observed in wild-type strains.Conclusions The molecular genetic characteristics of V-Oka strains are stable till the 50th passage.The results of sequence alignment show that pathogenic strains in chickenpox patients are wild-type strains,but not V-Oka strains.