水痘-带状疱疹病毒Oka株在人二倍体细胞株SV-1上的遗传稳定性及基因序列差异位点分析  

Genetic stability and nucleotide difference of varicella-zoster virus Oka strain subcultured in human diploid cell SV-1 strain

作　　者：孟凡红 于巍 张嵬[1,2] 杨利伟 何玉君 高强[1,2] 黄金凤[1,2] 尹卫东[1,2] 

机构地区：[1]科兴(大连)疫苗技术有限公司,辽宁大连116620 [2]北京科兴生物制品有限公司,北京100085

出　　处：《中国生物制品学杂志》2015年第2期121-128,共8页Chinese Journal of Biologicals

摘　　要：目的研究水痘-带状疱疹病毒（varicella-zoster virus,VZV）Oka株在人二倍体细胞株SV-1上的遗传稳定性,并对不同代次毒株进行全基因测序及差异位点分析。方法将VZV Oka株在SV-1细胞株上连续传至48代,检测病毒滴度。对VZV Oka株33、35、38、39及48代病毒进行全基因测序,应用DNASTAR.Lasergene.v 7.1软件进行数据的拼接及序列分析。结果 VZV Oka株在SV-1细胞上连续传代获得的33~48代病毒滴度在4.000~4.625 lg CCID50/ml之间,病毒滴度的算数平均值为4.292 lg CCID50/ml;VZV Oka株基因组全长125 114 bp,GC含量46%;33、35、38、39、48代病毒的核苷酸序列同源性为99.99%,这5代病毒与标准疫苗株V-Oka的核苷酸序列同源性为99.96%;VZV Oka株在MRC-5、SV-1细胞上制备的38代病毒全基因序列有1个差异位点;本研究中的各代次病毒基因序列高度同源,与Varivax和Varilrix疫苗株、野毒株Dumas及亲本株p-Oka相比,与疫苗株V-Oka的同源性最高。结论 VZV Oka株在SV-1细胞上的适应性及遗传稳定性均良好。Objective To investigate the genetic stability of varicella-zoster virus（VZV）Oka strain during subculture in human diploid cell SV-1 strain, and analyze the complete gene sequences and differential sites of Oka strain of various passages. Methods Oka strain was subcultured in SV-1 cells to passage 48, and determined for virus titer. The complete genes of passages 33, 35, 38, 39 and 48 were sequenced, of which the data were spliced and analyzed by using DNASTAR.Lasergene. v 7. 1 software. Results The virus titers of Oka strain of passages 33 ~ 48 were 4. 000 ~ 4. 625 lg CCID50/ ml,with an arithmetic mean of 4. 292 lg CCID50/ ml. The full-length of genome of Oka strain was 125 114 bp, while the GC content was 46%. The homology of nucleotide sequences of Oka strain of passages 33, 35, 38, 39 and 48 was 99. 99%,while that of the five passages to standard vaccine strain V-Oka was 99. 96%. One differential site was observed in the complete gene sequence of Oka strain subcultured to passage 38 in MRC-5 and SV-1 cells. High homologies were observed in the gene sequences of Oka strain of various passages. However, as compared with those of vaccine strains Vari-vax and Varilrix, wild strain Dumas and parent strain p-Oka, the homology of gene sequence of Oka strain to that of vac-cine strain V-Oka strain was high. Conclusion VZV Oka strain showed high adaptability and genetic stability in SV-1 cells.

关 键 词：水痘-带状疱疹病毒Oka株 遗传稳定性 差异位点 

分 类 号：R373.11[医药卫生—病原生物学] Q78[医药卫生—基础医学]