F0F1-ATPase生物传感器检测肠出血性大肠埃希菌O157:H7

Detection of Escherichia coli O157:H7 by F0F1-ATPase immune biosensors

中文关键词:  O157:H7  免疫生物传感器  检测

英文关键词:O157: H7  immune biosensor  detection

中文摘要:

      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(Escherichia coli)和沙门菌(Salmonella typhi)为干扰菌株进行了该方法的特异性研究。结果 检测时间为4.5 h, 102～104 cfu呈现良好的梯度性和线性, R2=0.9818。空白对照组和102 cfu/孔均与104 cfu/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。

英文摘要:

      Objective To develop a high sensitivity and specificity method for the detection of Escherichia coli O157:H7 by F0F1-ATPase immunobiosensors. Methods An immune biosensor based on the interference of load to the F0F1-ATPase rotation indicated by the fluorescence fluctuation was constructed to detect this pathogen. The concentration targets pathogen was measured according to the relative fluoresent value of F0F1-ATPase biosensor tagged by F1300 E.coli(ATCC14028) and Salmonella(CMCC44001) were tested as interferential bacteria to study the specificity of this method to O157: H7. Results The results dem-onstrated a good linearity (R2 = 0.9818) between antigen concentration (range from102 cfu to 104cfu) and the fluorescence intensity. The detection signals of the samples containing 102 cfu/well and 104 cfu/well O157: H7 were much stronger than the signal from control sample (P＜0.01). Conclusion Owing to its much higher sensibility and simplicity than other currently applied methods, it provides a promising future for its application in food source pathogen detection.