光纤生物传感器检测嗜肺军团菌方法的建立

摘要

Objective To establish the method of fiber optic biosensor for the detection of Legionella Pneumophila, in order to offer a fast, field detection mean for Legionella pneumophila. Methods At first, antigen or antibody was used to coat fibers; then the sample was added to incubate about 6 to 10 minutes; finally, the fluorescent probe was added to react 6 to 10 minutes. The purpose of qualitative or quantitative determination for the sample could be achieved, depending on the electrical signals converted from fluorescent signals by physicochemical transducer. Meanwhile, the effects of fiber modification methods, coated buffers and ionic strength on experimental results had been studied. Results When fiber optic biosensor used for Legionella pneumophila detection, its limit detection can achieve as low as 3x10 CFU/mL and its response time was about 20 min. The detection results of Salmonella Typhi, Escherichia Coli, Mycobacterium Tuberculosis, Yersinia Enterocolitica, Vibrio Cholerae, Staphylococcus Aureus and Shigella Dysenteriae were all negative, which showed favourable specificity. Conclusion Fiber optic biosensor shows advantages of rapid, high sensitivity, good specificity and reproducibility in detection of Legionella pneumophila. It can be used as a screening method for Legionella pneumophila detection.%目的 建立光纤生物传感器检测嗜肺军团菌的方法,为嗜肺军团菌提供一种快速、现场的检测手段.方法 先用抗原或抗体包被光纤,再加入待测物质孵育6～10 min,最后加入荧光探针反应6～10 min,根据理化换能元件可以将荧光信号转换成电信号的原理,实现对样品的定性或定量检测目的.同时探讨了光纤修饰方法,包被缓冲液以及离子强度等因素对实验结果的影响.结果 光纤生物传感器法检测嗜肺军团菌时,最低可检测到3×104 cfu/mL,检测响应时间为20 min；伤寒沙门菌、大肠杆菌、结核杆菌、小肠结肠炎耶尔森菌、霍乱弧菌、金黄色葡萄球菌、痢疾志贺菌的检测结果均为阴性,显示出良好的特异性.结论 光纤生物传感器检测嗜肺军团菌时,表现出快速、灵敏度高、特异性强、重复性好等优点,可作为嗜肺军团菌的一种初筛方法.