中国比较医学杂志2017，Vol.27Issue(9)：30-35,6.DOI:10.3969.j.issn.1671-7856.2017.09.006
转基因小鼠常见传染病血清学液相芯片技术多重检测方法的建立
Establishment of a flexible multi?analyte profiling assay to detect common infectious disease antibodies in the serum of transgenic mice
摘要
Abstract
Objective To establish a multiplex xMAP assay for detection of infectious disease antibodies in the serum of transgenic mice by liquid chips of mouse MHV, LCM, ECT and HANT proteins. Methods The fluorescent mi?crospheres were conjugated with mouse MHV, LCM, ECT and HANT proteins, and the amount of protein?conjugates and the working concentrations of antibodies and Streptavidin?PE were optimized, respectively. The liquid chips of mouse MHV, LCM, ECT and HANT were prepared and used to detect the antibodies in the serum samples from 56 transgenic mice, together with the negative, positive and control serum, by the multiplex xMAP assay. The results of xMAP assay were verified by traditional ELISA. Results (1) The optimal conjugated amount of mouse MHV, LCM, ECT and HANT protein was 20 μg, 20 μg, 40 μg and 20 μg, respectively. The optimal concentration of their detection antibodies was 2μg/mL, 2 μg/mL, 2 μg/mL and 4 μg/mL, and the optimal concentration of Streptavidin?PE for all of them was 2 μg/mL. (2) The results of xMAP assay showed that the MFI and index values of the serum samples from the No. 14, No. 23, No. 55 and No. 56 transgenic mice detected with the liquid chip of MHV were higher than those of the control serum, while other MFI and index values were lower than those of the control serum, indicating that the antibodies of MHV were positive in the No. 14, No. 23, No. 55 and No. 56 transgenic mice and negative in the other mice, and the antibodies of LCM, ECT and HANT were negative in all of the mice. (3) The results of ELISA showed that the A450 and index values of MHV in the No. 14, No. 23, No. 55 and No. 56 transgenic mice were higher than those of the control serum, while other A450 and index values were lower than those of the control serum, indicating that the antibodies of MHV were positive in the No. 14, No. 23, No. 55 and No. 56 transgenic mice and negative in the other mice, and the antibodies of LCM, ECT and HANT were negative in all of the mice. These ELISA results were consistent with the results of the xMAP assay. Conclusions The multiplex xMAP assay established in this study can be used in the detection of infectious disease antibodies in transgenic mice.关键词
转基因小鼠/传染病/xMAP技术/液相芯片Key words
Transgenic mice/Infectious diseases/Flexible multi⁃analyte profiling/xMAP/Liquid chip分类
医药卫生引用本文复制引用
陈诚,朱科燕,褚燕青,齐月寒,蔡月琴..转基因小鼠常见传染病血清学液相芯片技术多重检测方法的建立[J].中国比较医学杂志,2017,27(9):30-35,6.基金项目
浙江省科技计划项目(编号:2015C37111) （编号:2015C37111）
浙江中医药大学比较医学创新团队(编号:XTD201301). （编号:XTD201301）
