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目的研究F0F1-ATPase纳米生物传感器在肠出血性大肠埃希菌O157:H7检测中的应用,开发高灵敏、特异性强的快速检测技术。方法利用免疫技术构建信号识别元件,根据F0F1-ATPase酶学机制建信号转化系统,建立免疫生物传感器,最后通过荧光扫描仪进行信号检测。本研究以食品中常见的大肠埃希菌(Escherichiacoli)和沙门菌(Salmonellatyphi)为干扰菌株进行了该方法的特异性研究。结果检测时间为4.5h,102～104cfu呈现良好的梯度性和线性,R2=0.9818。空白对照组和102cfu/孔均与104cfu/孔组存在极显著性差异,P值分均为P=0.00<0.01。结论 F0F1-ATPase纳米生物传感器对O157:H7检测快速、灵敏、特异性好,有良好的应用前景。
Objective To study the application of F0F1-ATPase nano-biosensor in the detection of enterohemorrhagic Escherichia coli O157: H7 and to develop a sensitive and specific rapid detection technique. Methods Immunofluorescence was used to construct the signal recognition element. The F0F1-ATPase enzyme system was used to construct the signal transduction system to establish the immunobiosensor. Finally, the signal was detected by fluorescence scanner. In this study, Escherichia coli and Salmonella typhi, which are common in food, were used to study the specificity of this method. The results showed that the detection time was 4.5h, and 102 ~ 104cfu showed good gradient and linearity, R2 = 0.9818. There was a significant difference between the control group and 102cfu / well and the 104cfu / well group, P = 0.00 <0.01. Conclusion The F0F1-ATPase nanosensor has a rapid, sensitive and specific detection of O157: H7 and has good application prospects.