人用狂犬病疫苗株aGV的生物学特性  

Biological Characteristics of aGV Strain for Production of Rabies Vaccine for Human Use

作　　者：郭秀侠[1] 杨屹[1] 崔文广[1] 苗丽[1] 丁丽丽[1] 韩轼奇[1] 王亚军[1] 李宇辉[1] 

机构地区：[1]长春生物制品研究所有限责任公司,长春130062

出　　处：《中国生物制品学杂志》2011年第12期1460-1464,共5页Chinese Journal of Biologicals

摘　　要：目的研究人用狂犬病疫苗株aGV的生物学特性。方法取狂犬病病毒3aG株,接种于Vero细胞适应性传代,按照《中国药典》三部(2010版)要求建立毒种库,并进行全面检定:电镜观察毒种的形态结构,测定毒种的糖蛋白基因序列,直接免疫荧光法检测毒种的毒力,动物试验法和细胞培养法检测外源因子,免疫小鼠检测毒种的免疫原性,小鼠脑内中和试验法鉴定毒种的特异性,进行外周致病性试验,检测单次病毒收获液的效力。检测应用该毒株制备的疫苗的抗原性,并与上市疫苗进行比较。结果 aGV毒种在电镜下呈典型的子弹状,可见病毒包膜、刺突等结构;aGV株与3aG、PV、CTN-1、CVS株及街毒株糖蛋白基因序列的同源性分别为99%、92%、84%、91%、84%;病毒毒力逐代提高,第8～12代毒力稳定在108～109 FFU/ml;该病毒在Vero细胞上具有良好的适应性,外周致病性降低,无外源因子污染;aGV9和aGV12的中和指数分别为1 000和7 079,3个批次的aGV9免疫保护指数分别为38 019、7 943和13 489;中和抗体检测结果显示,试验组与对照组之间中和抗体效价差异无统计学意义(P>0.05)。结论本研究适应的狂犬病病毒aGV株有望作为人用狂犬病疫苗株应用于生产。Objective To investigate the biological characteristics of aGV strain for production of rabies vaccine for human use.Methods Rabies virus 3aG strain was inoculated to Vero cells for adaptive passage,based on which a virus seed bank was established according to the requirements in Chinese Pharmacopoeia（VolumeⅢ,2010 edition） and subjected to overall control tests.The virus seed was observed for morphology by electron microscopy,of which the glycoprotein gene was sequenced,then determined for virulence by direct IFA,for adventitious agents by animal test and cell culture method,for immunogenicity by immunization of mice,and for specificity by intracerebral neutralization test in mice.Peripheral pathogenicity test was performed on the virus seed,and the potency of a single virus harvest was determined.The immunogenicity of vaccine prepared with the strain was determined and compared with that of commercial vaccine.Results The aGV seed was in typical bullet-like shape under electron microscope,in which envelope and spikes were observed.The homologies of glycoprotein gene sequence of aGV strain were 99%,92%,84%,91% and 84% to those of 3aG,PV,CTN-1,CVS and street virus strain respectively.The virulence of virus strain increased with increasing passages,and was maintained at 108 ～ 109 FFU / ml from passages 8 to 12.The virus strain showed good adaptability to Vero cells,of which the peripheral pathogenicity decreased,and no adventitious agents were detected.The neutralization indexes of aGV9 and aGV12 were 1 000 and 7 079 respectively.The immune protection indexes of aGV9 of three batches were 38 019,7 943 and 13 489 respectively.No significant difference was observed between the neutralizing antibody titers induced by the prepared and commercial vaccines（P 0.05）.Conclusion The aGV strain adapted in this study might be used for production of rabies vaccine for human use.

关 键 词：狂犬病疫苗 aGV毒株 免疫原性 中和抗体 

分 类 号：R373.9[医药卫生—病原生物学] R392-33[医药卫生—基础医学]