防龋DNA疫苗免疫效能评价中间接ELISA检测的建立和验证  

Establishment and Verification of Indirect Enzyme-linked Immunosorbent Assay in Evaluating Immunity Effect of Anti-caries DNA Vaccine

作　　者：李宇红[1] 吴小丽[2] 闭兰[2] 郭长福[2] 张爱华[2] 

机构地区：[1]武汉大学口腔医学院口腔生物医学教育部重点实验室,湖北武汉430079 [2]武汉生物制品研究所

出　　处：《口腔医学研究》2011年第9期741-743,748,共4页Journal of Oral Science Research

基　　金：国家科技支撑计划资助项目(编号:2007BAI28B04);武汉市发展引导类计划资助项目(编号:201060938367)

摘　　要：目的:建立血清样本特异性抗体检测的间接ELISA方法,应用于龋齿DNA疫苗特异性抗体效力检测。方法:通过棋盘滴定确定样本检测条件建立血清特异性抗PAc抗体间接ELISA方法,并对所建立的方法进行特异性和精密度验证。结果:确定包被PAc蛋白浓度为10mg/L,酶标抗体工作浓度1∶5000,样本稀释倍数1∶200。方法的重复性检测表明,板间和板内误差小于15%;中间精密度高、中、低3个浓度的CV值小于15%。结论:该方法特异性强,具有良好的重复性和中间精密度,可用于防龋DNA疫苗免疫效能评价中血清样本特异性抗PAc抗体的检测。Objective： To evaluate the accuracy and reliability of an indirect enzyme-linked immunosorbent assay（ELISA） for detecting antibody to anti-caries DNA vaccine.Methods： Through the chessboard titration we grope the testing condition,and check the specific and precision of the methods.Results： The coating of PAc was 10μg/ml,the optimal dilution of the sera samples and goat anti-rat HRP labeled antibody was 1∶200 and 1∶5000,respectively.Repetitive test of methods indicated that measurement error was less than 15%.The coefficient of variation（CV） of intermediate precision high,medium and low concentrations was less than 15%.Conclusion： The indirect ELISA meets the requirements of repeatability and intermediate precision.It shows a potential application for antibody detection in anti-caries DNA vaccine as an alternative method.

关 键 词：防龋DNA疫苗 间接ELISA PAc蛋白 免疫 

分 类 号：R446.6[医药卫生—诊断学]