柯萨奇病毒A组16型多克隆抗体的制备及其在检定中的应用  

Preparation of polyclonal antibody against Coxsakie virus A16 and its application on virus vaccine test

作　　者：谢天宏[1] 杨婷[1] 宋霞[1] 李华[1] 岳磊[1] 刘正玲[1] 龙润乡[1] 杨蓉[1] 罗芳宇 朱凡丽 谢忠平[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所云南省重大传染病疫苗研究重点实验室,云南昆明650118

出　　处：《中国预防医学杂志》2017年第7期497-500,共4页Chinese Preventive Medicine

基　　金：云南省科技计划项目(2015BC008)

摘　　要：目的评价CV-A16-K168免疫血清对CV-A16不同毒株的中和抗体效价,确定完全中和不同病毒量所需的最低血清量,为CV-A16疫苗毒种检定中的鉴别试验和病毒外源因子检查提供参考。方法将CVA16病毒抗原辅以弗氏佐剂以皮下多点注射方式免疫新西兰大白兔,多次免疫后取血清,采用微量细胞病变法进行中和抗体效价检测和对15株不同CV-A16毒株的中和效价检测,并用CV-A16-4V和CV-A16-10R两个毒株分析中和不同量病毒所需的抗体量。结果浓缩抗原作为免疫原获得的兔抗CV-A16血清中和抗体效价可达1∶2 048;该免疫血清对15株CV-A16不同毒株的几何平均滴度(genomic mean titer,GMT)为1∶3 118;完全中和7.5lgCCID50/ml CV-A16病毒所需的最低血清量为128U,当病毒量在4.0~7.0lgCCID50/ml时,完全中和所需的最低血清量为48U,当病毒量低于4.0lgCCID50/ml时,1U的血清即可完全中和本病毒。结论 CV-A16-K168株作为免疫原可获得高效价的抗血清;用不同毒株检测得到的抗体效价存在差异;完全中和病毒所需的最低血清量与病毒量呈正相关;病毒量与测得的血清中和效价呈负相关。Objective To prepare the polyelonal antibody against Coxsakie virus A16 （CV-A16） and to evaluate its neutralization ability, determine the minimum amount of serum to neutralize different amount of CV-A16, and to provide reference for identification test for CV-A16 vaccine. Methods The New Zealand rabbits were subcutaneously immunized with the CV-A16 antigen in Freund＇s adjuvant and serum was collected after several boosts. Microdose eytopathogenie effect assay was used for the neutralization test of serum and the neutraliza- tion effect on 15 strains of CV-A16. CV-A16-4V and CV-A16-10R strains were then used to determine the minimum amount of serum used to neutralize different amount of virus. Results Neutralization titer of the anti-serum from rabbit immunized with concentrated CV-A16 antigen could reach to 1 ： 2 048. The genomic mean titer （GMT） of the anti-serum neutralizing 15 strains of CV-A16 was 1 ~ 3 118. It showed that the virus of 7.5 lgCCIDs0/ml was totally neutralized by 128 U serum, and 48 U serum could totally neutralize virus of 4.0-7.0 lgCCIDs0/mI. Only 1 U serum was required to totally neutralize virus when the amount of virus was lower than 4.0 tgCCIDs0/ml. Conclusions CV-A16-K168 virus can be used as a good immunogen to obtain the anti-serum with high neutralization ability. The titers of antiserum determined by different virus strains show some differences. The amount of virus used for titration is positively correlated with the minimum of ser um required to totally neutralize virus, while negatively correlated with the neutralization titer of anti serum.

关 键 词：柯萨奇病毒A组16型 免疫血清 中和能力 疫苗检定 

分 类 号：R392-33[医药卫生—免疫学]