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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 in human diploid cell line SV-1 and to analyze the whole genome and differential loci of different generations of strains . Methods VZV Oka strain was continuously passaged on SV-1 cell line for 48 passages to detect virus titer. Whole genome sequencing of VZV Oka strain 33, 35, 38, 39 and 48 generation viruses was carried out. DNASTAR.Lasergene.v7.1 software was used for data splicing and sequence analysis. Results The titers of 33-48 passages of VZV Oka strain were between 4.000 ~ 4.625 lg CCID50 / ml and the mean of virus titer was 4.292 lg CCID50 / ml. The VZV Oka strain genome The full-length cDNA was 125 114 bp with a GC content of 46%. The nucleotide sequence identities of 33, 35, 38, 39 and 48 were 99.99%. The nucleotide sequences of these 5-generation viruses and the standard vaccine strain V-Oka The homology was 99.96%. The VZV Oka strain had a different locus in the 38th generation of MRV-5 and SV-1 cells. The sequence of the virus in each generation in this study was highly homologous with Varivax had the highest homology with the vaccine strain V-Oka compared to the Varilrix vaccine strain, the wild-type Dumas strain and the parent strain p-Oka. Conclusion The adaptability and genetic stability of VZV Oka strain on SV-1 cells are good.