汉坦病毒感染的量子点标记免疫检测研究

目的 应用量子点荧光标记技术和间接免疫法,建立一种新的汉坦病毒（HV）感染的免疫检测技术.方法 采用碳二亚胺交联法,在水溶性量子点表面修饰protein G和羊抗人IgG.以HV抗原片为固相载体,QDs-PG-IgG复合物为标记二抗,检测待检血清中的HV特异性抗体,并对检测条件进行优化.结果 量子点与羊抗人IgG的最佳偶联条件：反应时间2h、pH 6.0、羊抗人IgG浓度20 μg/ml.检测体系的最佳反应条件：量子点标记二抗的最适工作浓度为1：200,HV感染阳性血清检测的最大稀释度为1：1280.结论 成功建立了简便、快速的HV感染的量子点标记免疫检测方法,该方法具有良好的灵敏度和特异性,抗荧光猝灭性强,为进一步实现HV抗体的单分子定量检测奠定基础.

A study of immunoassay by using quantum dots to detect Hantavirus infection

Objective To develop a new method to detect anti-Hantavirus IgG antibodies (HV IgG) based on quantum dots (QDs) and indirect immune technique. Methods The carbodiimide crosslinking method was used to couple protein G and goat antihuman IgG on the surface of water-solubility QDs.The coverglass covered HV antigen was used as carrier,and QDs-PG-IgG conjugates was used as labeled second antibody to detect the HV-IgG in the serum samples. The detecting conditions were optimized.Results The optimum reaction time,pH and goat antihuman IgG concentration for conjugating the QDs with goat antihuman IgG were 6.0,2h,and 20μg/ml,respectively.The optimum working dilution of QDs-PG-IgG conjugates was 1: 200.The detection limit of the serum samples was about 1:1280 dilution.Conclusion The method established in this study has been demonstrated to be a specific,sensitive,rapid test for detecting HV antibodies,laying the foundation of single molecule detection.The anti-fluorescence quenching ability of this method was significant improved.