An Immunobiosensor for Rapid Detection of Staphylococcus aureusenes
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摘要:
目的研究快速、经济、高灵敏的金黄色葡萄球菌新型检测技术。方法本研究应用F0 F1 -ATP 酶旋转分 子马达和免疫技术相结合，建立金黄色葡萄球菌的免疫生物传感器快速检测技术。首先pH 变化敏感的荧光物质 F1300 标记到F0 F1 -ATP 酶的载体色素体的内表面，然后在F0 F1 -ATP 酶上连接β 亚基抗体-生物素-链亲和素-生物 素-金葡菌抗体复合体，得到可以捕获金黄色葡萄球菌的免疫生物传感器。传感器上负载菌量不同，引起的酶活性 变化不同，酶活性变化又引起色素体内pH 变化，最后通过F1300 的荧光强度变化来检测菌量。结果该方法对金 黄色葡萄球菌标准菌株(CMCC26003) 的检测时间为4. 5 h，102 ～ 104 CFU /孔呈现良好的梯度关系。结论该技术 耗时短、操作简单、成本低、灵敏度高，符合食品安全检测技术发展要求。
Abstract:
Objective To develop a highly sensitive，low-cost and rapid assay method for the detection of Staphylococcus aureusenes. Method Combining F0 F1 -ATPase molecular motor with immunotechnology to establish a rapidly technology for detecting Staphylococcus aureusenes. F1300 is a fluorescence indicator sensitive to the change of pH. F1300 was labeled first on the inner surface of chromatophore，which is the carrier of F0 F1 -ATPase. A complex of Staphylococcus aureusenes antibody-biotin-streptavidin-biotin-β subunit monoclonal antibody was then connected to the β subunit of F0 F1 -ATPase to construct an immunobiosensor; then the Staphylococcus aureusenes can be captured by the antibody anchored on the complex. The activity of F0 F1 -ATPase changed with the content of Staphylococcus aureusenes loaded on the biosensor. The fluorescence of tagged F1300 was relative to the change of pH induced by the variation of F0 F1 -ATPase activities. Results The time taken by this examination was only 4. 5 h and a favorable gradient was shown at 102 － 104 CFU /well. Conclusion The method was time saving in examination，simple，low-cost and sensitive and met the requirements of detecting technology.
