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

发布时间：2018-03-02 11:26

本文选题：转基因小鼠　切入点：传染病　出处：《中国比较医学杂志》2017年09期 　论文类型：期刊论文

【摘要】：目的制备小鼠MHV、LCM、ECT和HANT液相芯片,建立转基因小鼠传染病抗体的xMAP多重检测方法。方法将荧光微球分别与小鼠MHV、LCM、ECT和HANT蛋白进行偶联,并且对最佳蛋白偶联量、检测抗体最佳工作浓度和Streptavidin-PE最佳工作浓度进行优化,制备小鼠MHV、LCM、ECT和HANT液相芯片,对56个转基因小鼠血清样品、阴性血清、阳性血清和质控血清进行xMAP多重检测。并应用传统的ELISA方法对xMAP检测结果进行验证。结果 1)小鼠MHV、LCM、ECT和HANT蛋白的最佳偶联量分别为20μg、20μg、40μg和20μg,检测抗体最佳浓度分别为2μg/mL、2μg/mL、2μg/mL和4μg/mL;Streptavidin-PE抗体最佳浓度均为2μg/mL;(2)xMAP检测结果显示,14号、23号、55号和56号转基因小鼠血清MHV MFI值和index值均高于质控血清,其余小鼠血清的MFI值和index值均低于质控血清,表明14号、23号、55号和56号转基因小鼠MHV抗体阳性,其余小鼠MHV、LCM、ECT和HANT抗体阴性;(3)ELISA检测结果显示,14号、23号、55号和56号转基因小鼠MHV A450值和index值均高于质控血清,其余小鼠血清的A450值和index值均低于质控血清,表明14号、23号、55号和56号转基因小鼠MHV抗体阳性,其余小鼠MHV、LCM、ECT和HANT抗体阴性,该ELISA检测结果与xMAP检测结果相一致。结论建立的xMAP多重检测技术可应用于转基因小鼠的传染病检测。
[Abstract]:Objective to prepare mouse MHVG LCMT and HANT microarray and to establish a xMAP multiplex assay for the detection of infectious disease antibodies in transgenic mice. Methods the fluorescent microspheres were coupled with mouse MHVLCMECT and HANT proteins respectively, and the optimal protein coupling amount was obtained. The best working concentration of antibody and the best working concentration of Streptavidin-PE were optimized to prepare mouse MHVN LCMECT and HANT liquid chip. 56 transgenic mice serum samples were obtained and negative serum samples were obtained. The results were verified by traditional ELISA method. Results 1) the optimal coupling amounts of MHVG LCMECT and HANT protein in mice were 20 渭 g, 20 渭 g and 20 渭 g respectively, and the optimal concentration of antibody was 20 渭 g. The best concentration of Streptavidin-PE antibody for 2 渭 g / mL and 4 渭 g / mL Streptavidin-PE antibody was 2 渭 g / mL / mL and 4 渭 g / mL respectively. The results showed that the serum MHV MFI and index values of transgenic mice of 14 渭 g / mLL, 55 渭 g / mL and 56 渭 g / mL were higher than those of QCS. The MFI and index values of the other mice serum were lower than those of the quality control serum, indicating that the MHV antibody was positive in the transgenic mice of 14, 23, 55 and 56. The results of Elisa showed that the MHV A450 and index values of the 14, 23, 55 and 56 transgenic mice were higher than those of the quality control serum, while the A450 and index values of the other mice were lower than those of the quality control serum. The results showed that the MHV antibodies were positive in transgenic mice of 14, 23, 55 and 56, while the other mice were negative for MHVG LCMECT and HANT antibodies. Conclusion the xMAP multiplex detection technique can be used in the detection of infectious diseases in transgenic mice.
【作者单位】：浙江中医药大学动物实验研究中心;
【基金】：浙江省科技计划项目(编号:2015C37111) 浙江中医药大学比较医学创新团队(编号:XTD201301)
【分类号】：Q95-33

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