乙型脑炎灭活疫苗生产用毒种P3-Smb53株全基因序列分析  

Complete genome sequencing of P3-Smb53 strain as virus seed for production of inactivated Japanese encephalitis vaccine

作　　者：刘欣玉[1] 俞永新[1] 徐宏山[1] 岳广智[1] 杨立宏[1] 董关木[1] 李玉华[1] 贾丽丽[1] 

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《中国生物制品学杂志》2014年第8期985-989,共5页Chinese Journal of Biologicals

摘　　要：目的 对我国乙型脑炎灭活疫苗生产用毒种P3株鼠脑53代（P3-Smb53）进行全基因序列测定及分析。方法提取P3-Smb53株病毒全基因组RNA,逆转录合成为cDNA,以其为模板分段进行PCR扩增,并测序;序列拼接后,利用NCBI的Blast程序和DNASTAR软件包中的MegAlign软件与GenBank中登录的5个基因型别的177株乙脑病毒全基因序列、551株乙脑病毒E蛋白序列进行比对。结果P3-Smb53株病毒基因组全长10 976个核苷酸,与177株乙脑病毒株的全基因序列核苷酸同源性为79.6%-99.7%,氨基酸同源性为91.3%-99.7%;与GenBank中登录的551株乙脑病毒E蛋白氨基酸同源性为91.0%-99.8%,在E蛋白12个关键抗原性位点中,所有毒株均非常保守;与GenBank中登录的P3株全基因序列核苷酸同源性为99.4%,氨基酸同源性为99.5%。结论 乙型脑炎灭活疫苗生产用毒种P3-Smb53株虽然与不同基因型毒株间的氨基酸差异较大,但在关键抗原位点上高度一致,理论上能够抵抗目前国内外所有乙脑分离株的感染。Objective To determine and analyze the complete genome sequence of P3 strain subeultured in mouse brain for 53 passages （P3-Smb53） used for production of inactivated Japanese encephalitis （JE） vaccine. Methods Complete genomic RNA of P3-Smb53 strain was extracted and reversely transcribed to eDNA which was used as a template for fractional amplification by PCR. The amplified seven fragments were sequenced and spliced, and the results were compared with the complete genome sequences of 177 JE virus strains of 5 genotyes and the E protein sequences of 551 JE virus strains in GenBank by using the Blast program in NCBI and MegAlign software in DNASTAR software package. Results The genome of P3-Smb53 strain was 10 976 nueleotides in length, of which the homologies of nucleotides and amino acids to those of 177 JE virus strains in GenBank were 79. 6% = 99. 7% and 91. 3% =99. 7% respectively. However, the homology of amino acids of P3-Smb53 strain was 91.0% = 99. 8% to that of E protein of 551 JE strains in GenBank. All the 12 critical antigenic epitopes were highly conserved in various strains. The homologies of nucleotides and amino acids of P3-Smb53 strain to those of P3 strain in GenBank were 99.4% and 99. 5% respectively. Conclusion The amino acid sequence of P3-Smb53 strain was significantly different from, while the critical antigenic epitopes were highly consistent with, those of other strains of different genotypes, indicating that the inactivated JE vaccine prepared with P3-Smb53 strain might offer protection against the infection with all the JE virus strains isolated home and abroad.

关 键 词：流行性乙型脑炎 灭活疫苗 毒种 序列分析 

分 类 号：R373.31[医药卫生—病原生物学] Q789[医药卫生—基础医学]