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目的建立一种无需增菌即能快速、灵敏检测大肠杆菌O157:H7的新方法。方法制备一种表面包被大肠杆菌O157:H7单克隆抗体的纳米级免疫磁颗粒,用于样品中菌体的富集和分离;制备一种表面分别标记有大肠杆菌O157:H7多克隆抗体和生物素化核酸探针的胶体金颗粒;利用生物素-亲和素-酶系统信号放大作用,达到对目标菌的快速、灵敏检测。结果该方法可在1h内完成菌体的分离和检测,检测限为10cfu/ml。结论该检测体系基于功能化纳米粒子进行菌体分离及信号放大,具有检测时间短、灵敏度高、操作简便等优点,在环境、食品检测,临床诊断和流行病监测中具有广泛的应用前景。
Objective To establish a new method for rapid and sensitive detection of Escherichia coli O157: H7 without increasing bacteria. Methods A nano-immunomagnetic particle coated with the surface-coated E. coli O157: H7 monoclonal antibody was prepared for the enrichment and separation of the bacterial cells in the sample. A polyclonal antibody labeled with Escherichia coli O157: H7 Biotinylated nucleic acid probe colloidal gold particles; the use of biotin - avidin - enzyme system signal amplification, to achieve rapid and sensitive detection of the target bacteria. Results The method can be completed within 1h isolation and detection of bacteria, detection limit of 10cfu / ml. Conclusion The detection system based on functionalized nanoparticles for cell separation and signal amplification has the advantages of short detection time, high sensitivity and easy operation, and has broad application prospects in environment, food testing, clinical diagnosis and epidemiological surveillance.