Abstract：
Rapid, sensitive and simultaneous detection of multiple bacteria in foodborne is still a major challenge in public health field. Here, a fluorescence immunoassay that can achieve high-throughput detection of three Gram-positive foodborne pathogenic bacteria simultaneously was proposed. Vancomycin and bovine serum albumin conjugate (Van-BSA) was immobilized on a polycarbonate chip to capture three Gram-positive foodborne pathogenic bacteria, Staphylococcus aureus (S. aureus), Bacillus cereus (B. cereus) and Listeria monocytogenes (L. monocytogenes). CdSe/ZnS quantum dot modified antibodies (Ab-QD) were prepared by carbodiimide coupling chemistry. Due to the affinity reaction between antibodies and proteins on the bacterial surface, the simultaneous detection of multiple Gram-positive bacteria was achieved by monitoring the fluorescence signal of quantum dot by a portable microfluidic chip analyzer. Under optimal conditions, low detection limits was 18 CFU/well, 3 CFU/well and 36 CFU/well for S. aureus, B. cereus and L. monocytogenes, respectively. With satisfactory accuracy and precision, the proposed fluorescence immunoassay holds good prospects to detect pathogens in real food samples.
摘要：
快速，同时检测食源性多种细菌仍然是公共卫生领域面临的重大挑战。这里,提出了一种能同时实现3种革兰氏阳性食源性致病菌高通量检测的荧光免疫分析方法。万古霉素和牛血清白蛋白缀合物（Van-BSA）固定在聚碳酸酯芯片上，以捕获三种革兰氏阳性食源性致病菌，金黄色葡萄球菌（S。金黄色葡萄球菌)，蜡样芽孢杆菌(B.蜡状细胞）和单核细胞增生李斯特菌（L.单核细胞增多症）。采用碳二亚胺偶联化学方法制备了CdSe/ZnS量子点修饰抗体(Ab-QD)。由于抗体和细菌表面的蛋白质之间的亲和反应，通过便携式微流控芯片分析仪监测量子点的荧光信号,实现了多种革兰氏阳性菌的同时检测。在最优条件下,低检测限为18CFU/孔，金黄色葡萄球菌3CFU/孔和36CFU/孔，蜡状芽孢杆菌和单核细胞增生李斯特菌，分别。具有令人满意的准确度和精密度，所提出的荧光免疫分析法在实际食品样品中检测病原体具有良好的前景。
