水痘-带状疱疹病毒抗血清的制备及检定  

Preparation and quality control of antiserum against varicella-zoster virus

作　　者：刘洪涛 孙跃秋 王冲 王梦涵 张树杰 李航 庞惠娟 徐俊峰 张夫坤 LIU Hongtao;SUN Yueqiu;WANG Chong;WANG Menghan;ZHANG Shujie;LI Hang;PANG Huijuan;XU Junfeng;ZHANG Fukun(Changchun Keygen Biological Products Co.Ltd.,Changchun 130062,Jilin Province,China)

机构地区：[1]长春祈健生物制品有限公司研究室,吉林长春130062

出　　处：《中国生物制品学杂志》2024年第8期983-988,共6页Chinese Journal of Biologicals

基　　金：吉林省科技发展计划项目(20200404191YY)。

摘　　要：目的制备水痘-带状疱疹病毒(varicella-zoster virus,VZV)收获物,纯化后免疫日本大耳白兔,制备VZV抗血清,并进行各项检定。方法将VZV Oka株(vOka)在人二倍体细胞2BS株中传代培养,病毒收获后经超速离心纯化,制备免疫原,与弗式佐剂混合后经背部多点注射免疫雌性日本大耳白兔5只,制备VZV抗血清。Western blot检测血清抗体的特异性;测定膜抗原荧光抗体(fluorescent antibody to membrane antigen,FAMA)效价和血清抗体中和效价,并对两种检测方法的结果进行相关性分析。利用抗体效价较高的血清对麻疹、腮腺炎和风疹病毒进行异种病毒干扰试验。结果制备的VZV抗血清可特异性结合vOka和Oka-7S病毒收获物及gE和ORF7蛋白等多种VZV抗原;3免血清FAMA抗体效价可达1∶16384,血清抗体中和效价达1∶256;根据Karl Pearson相关系数分析得出两种检测方法结果呈线性相关。异种病毒干扰试验中试验组与对照组病毒滴度差值均<0.50 lgCCID50/mL,表明VZV抗血清对麻疹、腮腺炎和风疹病毒滴度均无干扰。结论成功制备了特异性强,亲和力高的VZV抗血清,为VZV成分疫苗的检定奠定了基础。Objective To prepare varicella-zoster virus(VzV)harvest and immunize Japanese rabbits with purified bulk of virus to prepare antiserum against VZV,and to perform various controltests.Methods The VZV Oka strain(vOka)was subcultured in human diploid cell line 2BS.The harvested virus was purified by ultracentrifugation to prepare immunogen,which was mixed with Freund's adjuvant and then used to immunize 5 female Japanese white rabbits by multi-point subcuta-neous injection to prepare VZV antiserum.The specificity of serum antibody was detected by Western blot.The titers of fluo-rescent antibody to membrane antigen(FAMA)and serum neutralizing antibody were determined,and the correlation between the results of the two detection methods was analyzed.Interference of heterogenous viruses was performed on measles,mumps and rubella viruses by using antiserum with high antibody titer.Results The prepared VZV antiserum had specific binding with vOka and Oka-7S virus harvests,gE and ORF7 proteins and other VZV antigens.After the third immu-nization,the serum antibody titer detected by FAMA was 1:16384,and the neutralizing antibody titer was 1:256.According to Karl Pearson correlation coefficient analysis,the results of the two detection methods were linearly corre-lated.Interference of heterogenous viruses showed that the differences between the experimental and control groups were less than O.50 IgCCIDso/mL,indicating that the antisera against VZV had no interference to the titers of measles,mumps and rubella viruses.Conclusion The VZV antiserum with high specificity and affinity was successfully prepared,which lays a foundation for the validation of VZV component vaccines.

关 键 词：水痘-带状疱疹病毒 中和抗体 兔抗血清 

分 类 号：R373.9[医药卫生—病原生物学]