CAJ | 학술논문

目的研究流行性乙型脑炎Vero细胞灭活疫苗毒种(P3毒株)在生产过程中的遗传稳定性,为疫苗的安全性和免疫原性评价提供依据.方法检测P3毒株鼠脑传代一代毒株、主种子、工作种子、疫苗及疫苗续传5代后病毒E蛋白基因核苷酸及氨基酸序列,并与GenBank中乙脑病毒P3株(AF036919)进行比对分析,同时比较主种子、工作种子、疫苗及疫苗续传5代后毒株的病毒滴度、抗原含量及效价.结果以上5代次病毒的E蛋白基因核苷酸及蛋白质序列完全相同,同源性均为100％,与GenBank中乙脑病毒P3株(AF036919)比较显示:核苷酸序列中第E9、E10、E324、E330、E1223、E1338基因位点存在差异,同源性为99.73％,其中只有E1223突变引起相应氨基酸aE408突变(L→S),同源性为99.80％,但该位点为非毒力相关位点,而其他位点均为沉默突变.4个代次疫苗毒株病毒滴度均≥8.0 lgLD50/ml,且各代次抗原含量及效价较为相似.结论在生产乙型脑炎灭活疫苗(P3株)过程中建立的以Vero细胞为基质的乙脑毒种库具有良好的遗传稳定性.
목적 연구류행성을형뇌염Vero세포멸활역묘독충(P3독주)재생산과정중적유전은정성,위역묘적안전성화면역원성평개제공의거.방법 검측P3독주서뇌전대일대독주、주충자、공작충자、역묘급역묘속전5대후병독E단백기인핵감산급안기산서렬,병여GenBank중을뇌병독P3주(AF036919)진행비대분석,동시비교주충자、공작충자、역묘급역묘속전5대후독주적병독적도、항원함량급효개.결과 이상5대차병독적E단백기인핵감산급단백질서렬완전상동,동원성균위100％,여GenBank중을뇌병독P3주(AF036919)비교현시:핵감산서렬중제E9、E10、E324、E330、E1223、E1338기인위점존재차이,동원성위99.73％,기중지유E1223돌변인기상응안기산aE408돌변(L→S),동원성위99.80％,단해위점위비독력상관위점,이기타위점균위침묵돌변.4개대차역묘독주병독적도균≥8.0 lgLD50/ml,차각대차항원함량급효개교위상사.결론 재생산을형뇌염멸활역묘(P3주)과정중건립적이Vero세포위기질적을뇌독충고구유량호적유전은정성.
Objective To investigate genetic stability of P3 strain of Vero cell derived inactivated Japanese encephalitis vaccine.Methods The nucleotide and amino acid sequences of E protein of Japanese encephalitis virus (JEV) P3 strain at different culture period including the mouse brain one passage seed,master seed,working seed the vaccine lot in addition to its 5 passages lot were determined,while the E gene and protein sequences were compared with JEV wide stain (AF036919) from the GenBank.Furthermore,the master seed,working seed,vaccine lot and its 5 passages lot of P3 stain were determined for virus titer,antigen concentration and the vaccine potency.Results The E gene and protein sequences of the above 5 passages of vaccine strain prepared for JEV vaccine showed no difference with homologies of 100％.When the 5 passages of vaccine stain compared with those JEV wide stain (AF036919),the gene sequence at E9,E10,E324,E330,E1223,E1338 showed difference with homologies of 99.73％.No silent mutation were investigated except the amino acid mutation at aE408 (L→S) but was no virulence-associated sites with homologies of 99.80％.The virus titers of the master seed,working seed,vaccine lot and its 5 passages lot of P3 stain were higher than 8.0 lgLD50/ml,while the antigen concentrations and the vaccine potency showed no difference.Conclusion The virus seed bank of P3 strain for Vero cell derived inactivated Japanese encephalitis vaccine showed high genetic stable.