人乳头瘤病毒假病毒中和试验血清起始稀释倍数的研究  

Investigation of initial serum dilution in pseudovirion neutralization antibody assay for detecting antibodies against human papillomavirus

作　　者：赵慧[1] 李娟[1] 马新兴 周凌云 李长贵[1] ZHAO Hui;LI Juan;MA Xin-xing;ZHOU Ling-yun;LI Chang-gui(National Institutes for Food and Drug Control, Division of Respiratory Virus Vaccines, Beijing 100050, China)

机构地区：[1]中国食品药品检定研究院呼吸道病毒疫苗室卫生部生物技术产品检定方法及其标准化重点实验室,北京100050 [2]上海泽润生物科技有限公司研发室,上海201203

出　　处：《微生物学免疫学进展》2018年第2期1-4,共4页Progress In Microbiology and Immunology

基　　金：国家国际科技合作专项(2013DFA31680)

摘　　要：目的对人乳头瘤病毒(human papilloma virus,HPV)假病毒中和试验起始稀释倍数进行研究,确定该系统中合适的起始稀释倍数。方法为了排除血清中HPV抗体以及母传抗体的干扰,选取50份15~18月龄的儿童血清作为验证样本。血清以不同的稀释倍数,与适量的假病毒中和。以光学显微镜观察接种细胞的形态,来判定不同稀释度的血清对细胞生长状况的影响;观察感染细胞表达的绿色荧光蛋白的强度及数量,以此考察血清对假病毒感染过程的影响。结果血清稀释度1∶40对细胞生长状况影响较小;该稀释倍数的病毒进入细胞表达的荧光强度与不加血清的阳性对照类似;假病毒感染细胞导致的荧光斑点数与真实病毒对照相比减少了10%,远低于中和试验判定阈值(相对抑制率50%)。结论在该实验系统中,HPV假病毒中和试验的血清起始稀释度以1∶40较为合适。Objective To investigate an initial serum dilution in the pseudovirion-based neutralization assay （PBNS） for detecting antibodies against human papillomavirus （HPV）. Methods To remove the interference from the pre-existing HPV antibodies and maternal transfected antibodies, a total of 50 serum samples were taken as validated samples from chil- dren aged 15-18 months in this study. The different dilutions of serum were neutralized with the corresponding mount of HPV. The inoculated cell morphology were observed under the light microscope to judge the influence from the different concentration of serum on the cell growth ; the strength and the amount of green fluorescent protein produced from the infec- ted cells were observed to learn the effect of serum on the process that HPV entered the cells. Results The serum with a 1 ： 40 dilution had less effect on the cell morphology; under the same condition, the fluorescence intensity expressed by in- fected cells with pseudovirion was similar to that of the positive control, fluorescent spots decreased 10% than the positive virus control, far lower than the criterion of 50% in the PBNS. Conclusion In this experiment system, The serum with a 1 ： 40 dilution as the initial concentration was suitable in PBNS for detecting antibodies against HPV.

关 键 词：人乳头瘤病毒 中和试验 假病毒 方法 验证 

分 类 号：R373[医药卫生—病原生物学]