【摘要】 建立了一种基于表面等离子体共振(surface plasmon resonance,SPR)原理的生物传感器方法,实现了快速检测大肠杆菌O157∶H7。研究选用BIACORE 3000系统及葡聚糖修饰的CM5芯片,先用EDC/NHS将芯片活化,然后抗体直接通过酰胺键固定在金表面,再用乙醇胺封闭,这样处理之后的芯片就可用于检测大肠杆菌O157∶H7。利用NaOH溶液对芯片再生,实现对多个不同浓度样品检测,采用时间对响应单位(RU)记录数据。该法检测大肠杆菌O157∶H7的检测限为3×105CFU.mL-1,RU变化值和大肠杆菌O157∶H7的浓度在一定范围内相关性良好,相关系数达到0.99。检测时间短,一个样品仅需5~7 min,再生效果好,芯片可重复使用50次以上。可快速、在线、稳定地检测大肠杆菌O157∶H7,有望成为一种在线检测食品致病性微生物的有力手段。更多还原

【Abstract】 A biosensor based on surface plasmon resonance(SPR) was developed for rapid detection of Escherichia coli O157∶H7.BIACORE 3000 SPR instrument and a dextran-modified sensor chip(CM5) were used.After activation with EDC/NHS,anti-E.coli O157∶H7 antibody was immobilized on the gold surface of the SPR sensor,and then following ethanolamine was injected,and the chip was ready for E.coli O157∶H7.Regeneration was achieved using NaOH in order to detect several samples.The limit of detection was found to be 3×105 CFU·mL-1 for E.coli O157∶H7,and the change of RU was linearly correlated with the concentration of E.coli O157∶H7 cells(R2=0.99).The detection time ranged from 5 to 7 min,and the result of regeneration was effective which allowed the chip to be reused for more than 50 samples.This method is convenient,and stable,and shows potentials for applications in food areas.更多还原