摘要
目的建立并验证重组人乳头瘤病毒双价(16/18型)疫苗中和滴度的检测方法 ,并且利用此假病毒法进行免疫原性研究。方法建立并优化以ZsGreen假病毒为基础的中和实验(ZsGreen法)测定血清中和滴度,比较荧光显微镜观察与微流式细胞分析法、流式细胞分析法的差异,并对专属性、精密度、耐用性、假病毒稳定性和不同细胞代次的影响进行了方法学验证。结果ZsGreen法的HPV16/18假病毒最佳接种量为1 600 TCID50,HPV16型与HPV18型无交叉,具有良好的专属性和精密度,采用不同细胞代次与不同批次假病毒进行检测,均能获得较好的重复性。结论 ZsGreen法假病毒中和滴度检测法准确可靠、重复性好,经方法学验证后此法适用于疫苗的免疫原性研究。
This study designed to establish and validate a neutralizing antibody titer assay for recombinant human papillomavirus（16/18） candidate vaccine, and then to evaluate the immunogenicity by this method. Based on pseudovirus, ZsGreen neutralizing antibody titer assay was optimized. And then, we compared the result of fluorescence microscopy with that of personal flow cytometry and flow cytometry. At last, the specificity, precision,robustness, and stability of pseudovirus assay and the cell generation time were validated. Data showed that the optimized concentration of HPV pseudovirus（types 16 and 18） for ZsGreen was 1 600 TCID50, and HPV16 did not crossreacted with HPV18. The assay also showed good specificity and precision. Furthermore, the assay demonstrated good repeatability, when different generation cell and different lots of pseudovirus were used. In conclusion, the constructed ZsGreen pseudovirus neutralizing antibody titer assay possesses advantages of repeatability and precision, thus adapts to evaluate immunogenicity of vaccine.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2014年第8期731-736,共6页
Immunological Journal
基金
国家"十二五"重大新药创制(2011ZX09102-001-22)
上海市科学技术委员会项目(12431901900)
