水痘减毒活疫苗Oka株的传代稳定性  

Stability of Live Attenuated Varicella Vaccine Oka Strain during Subculture

作　　者：陈哲文[1] 金于兰[1] 瞿爱东[1] 马相虎[1] 王亮[1] 杨忠东[1] 

机构地区：[1]上海生物制品研究所,上海200052

出　　处：《中国生物制品学杂志》2010年第12期1333-1337,共5页Chinese Journal of Biologicals

基　　金："十一五"国家科技支撑计划重点项目(2008BAI54B03)

摘　　要：目的观察水痘减毒活疫苗Oka株(V-Oka)的传代稳定性。方法将V-Oka病毒在人二倍体细胞MRC-5上连续传代至50代,观察病毒的培养特性。选择32、34、38、42、46、50代病毒进行病毒滴度测定和鉴别试验,提取病毒DNA,对V-Oka的主要基因(ORF31、ORF62、ORF68、R区)进行PCR扩增和测序,并对序列进行分析。结果 V-Oka连续传代至50代,其培养特性、细胞病变一致,病毒滴度为5.0～5.6LgPFU/ml。与GenBank中登录的V-Oka序列比较,32～50代的V-OkaORF31、ORF62、ORF68、R区序列稳定,并趋于减毒型。结论 V-Oka传代至50代,生物学和分子遗传学特性稳定。Objective To observe the stability of live attenuated varicella vaccine Oka strain（V-Oka）during subculture.Methods V-Oka strain was subcultured in human diploid MRC-5 cells to passage 50 and observed for cultural characteristics.The virus of passages 32,34,38,42,46 and 50 were selected for titration and identity test.The viral DNAs were extracted,from which the major genes（ORF31,ORF62,ORF68 and R regions）were amplified by PCR and sequenced.Results After subculture for 50 passages,the cultural characteristics and CPEs of V-Oka strain showed no significant change,and the virus titer was 5.0 ～ 5.6 LgPFU/ml.Compared with those reported in GenBank,the sequences of ORF31,ORF62,ORF68 and R regions were stable.With the increasing passages,the strain showed an attenuated tendency.Conclusions The biological and molecular genetic characteristics of V-Oka strain were stable after subculture to passage 50.

关 键 词：水痘疫苗 疫苗 减毒 Oka株 连续传代 稳定性 

分 类 号：R373.1[医药卫生—病原生物学]