基于定向固定抗体Fab片段的免疫层析技术同时检测中药中多种真菌毒素的研究

Mycotoxins are a class of typical harmful substances in traditional Chinese medicine. The mycotoxins are monitored and analyzed in traditional Chinese medicine, and this is one of the important contents for the quality and safety of traditional Chinese medicine. To establish a simple, rapid and reliable immunechromatography technology for screening and detection of multiple mycotoxins in traditional Chinese medicine is becoming more and more important. In this study, the immunochromatographic strips of simultaneous detection for two or three kinds of mycotoxins will be prepared by the connection with oriented immobilization of antibody Fab fragment and urchin-like gold nanoparticle. The preparation conditions will be optimized. The sensitivity, stability, specificity and repeatability of the test strip will be evaluated, and the evaluation method will be developed by using the test strip for detection mycotoxins in traditional Chinese medicine. The immunochromatographic strips will be used for rapid screening detection of mycotoxins in traditional Chinese medicine for the health supervision, pharmaceutical companies, hospitals and family. The results of the project will be provide technical support for effective prevention and control of mycotoxin contamination in the traditional Chinese medicine, and have good social and economic value.

真菌毒素是中药中存在的一类典型的有害物质，加强对中药中的真菌毒素监控和分析，是中药质量安全评价的重要内容之一。建立快速、简单和可靠的同时对中药中多种真菌毒素的筛查和检测的免疫层析技术显得越来越重要。本项目拟通过将抗体的Fab片段定向固定后，再与金纳米花颗粒结合，优化各项工艺条件，制备出同时检测两种和三种真菌毒素的高灵敏度免疫层析试纸条。同时对试纸条的灵敏度、稳定性、特异性和重复性等性能进行评价，并开展其在中药中真菌毒素检测中的应用评价研究。项目的研究成果可应用于卫生监督、药厂、医院和家庭等现场快速初筛检测，从而为有效预防和控制中药中真菌毒素污染提供技术支撑，具有良好的社会和经济价值。

真菌毒素是中药中存在的一类典型的有害物质，加强对中药中的真菌毒素监控和分析，是中药质量安全评价的重要内容之一。本项目对真菌毒素（AFB1、DON、ZEN、FB1、OTA和T-2 等）抗体进行了的纯化，并对真菌毒素抗体的Fab片段进行了酶解；分别对球形胶体金和金纳米花的制备条件和试纸条的制备条件进行了优化，分别制备了单独检测（AFB1、DON、ZEN、FB1、OTA和T-2）和同时检测FB1和DON；同时检测AFB1、ZEN和T-2；同时AFB1、ZEN和OTA；同时检测DON、ZEN和OTA球形和金纳米花型免疫层析试纸条；同时考察了胶体金和金纳米花颗粒形貌和大小对免疫层析试纸条性能的影响；对制备的试纸条灵敏度、特异性和稳定性等进行了评价；采用制备的单独检测和同时检测多种真菌毒素试纸条对实际中药样品进行了检测，然后采用HPLC和ELISA方法对试纸条的检测结果进行了对照评价，确定了试纸条检测方法的准确性。项目的研究成果为有效预防和控制中药中真菌毒素污染提供技术支撑，具有良好的社会和经济价值。