利用DNA免疫技术制备麻疹病毒抗血清  

Preparation of antiserum against measles virus by using DNA immunization technology

作　　者：权娅茹[1] 崔晓雨[1] 易敏[1] 徐涵 陈震[1] 邱平[1] 李长贵[1] 袁力勇[1] 

机构地区：[1]中国食品药品检定研究院,北京100050 [2]苏州杰恩生物技术有限公司,江苏苏州215123

出　　处：《中国生物制品学杂志》2016年第9期984-986,共3页Chinese Journal of Biologicals

摘　　要：目的利用DNA免疫技术制备特异性麻疹病毒抗血清,用于麻疹减毒活疫苗毒种以及含麻疹病毒疫苗的检定。方法以含麻疹病毒沪-191融合蛋白F和血凝素H基因的重组真核表达载体pGI-F和pGI-H作为免疫原,免疫家兔,制备麻疹病毒抗血清。采用细胞培养法检测抗血清细胞毒性,ELISA法检测抗体滴度,微量细胞病变法检测抗血清的特异性和中和效价。结果麻疹病毒抗血清细胞毒性小,与风疹病毒、腮腺炎病毒、水痘病毒均无交叉反应。抗血清的ELISA效价大于256 000,中和效价为1∶1 024,4倍稀释后仍能完全中和1×10 5 CCID50/ml麻疹病毒。结论 DNA免疫方法可制备高效价、低细胞毒性的麻疹病毒抗血清,该血清可用于麻疹疫苗毒种及含麻疹疫苗的质量控制。Objective To p repare specific antiserum against measles virus by using DNA immunization technique and use for the control tests on live attenuated measles vaccine virus seed and measles-containing vaccine. Methods Rabbits were immunized with recombinant plasmids p GI-F and p GI-H, containing the fusion protein F and hemagglutinin H genes of Hu-191 strain, and the prepared antiserum were determined for cytotoxicity by cell culture method, for antibody titer by ELISA, and for specificity and neutralizing titer by micro-CPE method. Results The antiserum against measles virus showed low cytotoxicity, while no cross-reaction with rubella virus, mumps virus or varicella virus. The ELISA titer of the antiserum was more than 256 000, while the neutralizing titer was 1 ∶ 1 024. The antiserum diluted 4-fold completely neutralized 1 × 10 5 CCID50/ ml measles virus. Conclusion Antiserum against measles virus, with high titer and low cytotoxicity, were prepared by DNA immunization method, which might be used for the quality control of measles vaccine virus seed and measles-containing vaccine.

关 键 词：DNA免疫 家兔 麻疹病毒 抗血清 中和效价 

分 类 号：R373.11[医药卫生—病原生物学] R392-33[医药卫生—基础医学]