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目的建立基于量子点和免疫磁珠技术检测沙门菌、志贺菌和金黄色葡萄球菌的方法。方法利用水相法合成多色的CdSe量子点,通过共价交联法将三色量子点与抗沙门菌、抗志贺菌和抗金黄色葡萄球菌的特异性抗体偶联,利用紫外吸收光谱和SDS-PAGE电泳证明偶联是否成功。利用免疫磁珠富集,量子点标记,通过荧光显微成像法对沙门菌、志贺菌和金黄色葡萄球菌进行检测。结果多色量子点与3种菌的特异性抗体偶联成功,通过选用3种不同颜色不同发射波长的量子点作标记,利用免疫磁珠富集3种菌,在同一反应体系中同时检测3种菌,检测时间在2h之内。对模拟感染目标菌的样品直接检测,检测限为103CFU/ml。结论该方法能够快速、特异性地检测沙门菌、志贺菌和金黄色葡萄球菌。
Objective To establish a method for detecting Salmonella, Shigella and Staphylococcus aureus based on quantum dots and immunomagnetic beads technology. Methods The multicolor CdSe QDs were synthesized by aqueous phase method. Three color QDs were conjugated with specific antibodies against Salmonella, Shigella and Staphylococcus aureus by covalent cross-linking method. The UV absorption spectra And SDS-PAGE electrophoresis to prove the success of the coupling. Salmonella, Shigella and Staphylococcus aureus were detected by fluorescence microscopy using immunomagnetic beads enrichment and quantum dot labeling. Results The coupling of multi-color quantum dots with the specific antibodies of three kinds of bacteria was successful. Three quantum dots with different emission wavelengths were selected for labeling. The immunomagnetic beads were used to enrich three kinds of bacteria and simultaneously detected in the same reaction system. Inoculum, detection time within 2h. Direct detection of samples simulating infection of target bacteria, detection limit of 103CFU / ml. Conclusion The method can detect Salmonella, Shigella and Staphylococcus aureus rapidly and specifically.