Department of Food Science and Human Nutrition, The University of Maine, 5735 Hitchner Hall, Orono, Maine 04473, USA.
Biosens Bioelectron. 2012 Oct-Dec;38(1):177-83. doi: 10.1016/j.bios.2012.05.024. Epub 2012 May 29.
A sensitive bacteria enrichment and detection system for viable Escherichia coli O157:H7 was developed using a piezoelectric biosensor-quartz crystal microbalance (QCM) with antibody-functionalized gold nanoparticles (AuNPs) used as detection verifiers and amplifiers. In the circulating-flow QCM system, capture antibodies for E. coli O157:H7 were first immobilized onto the QCM chip. The sample containing E. coli O157:H7 was circulated through the system in the presence of 10 ml of brain heart infusion (BHI) broth for 18 h. The cells of E. coli O157:H7 specifically captured and enriched on the chip surface of the QCM were identified by QCM frequency changes. Listeria monocytogenes and Salmonella Typhimurium were used as negative controls. After bacterial enrichment, detection antibody-functionalized AuNPs were added to enhance the changes in detection signal. The use of BHI enrichment further enhanced the sensitivity of the developed system, achieving a detection limit of 0-1 log CFU/ml or g. The real-time monitoring method for viable E. coli O157:H7 developed in this study can be used to enrich and detect viable cells simultaneously within 24h. The unique advantages of the system developed offer great potential in the microbial analysis of food samples in routine settings.
采用抗体功能化金纳米粒子(AuNPs)作为检测验证和放大的压电生物传感器-石英晶体微天平(QCM),开发了一种用于检测活大肠杆菌 O157:H7 的敏感细菌富集和检测系统。在循环流动 QCM 系统中,首先将大肠杆菌 O157:H7 的捕获抗体固定在 QCM 芯片上。在存在 10ml 脑心浸液(BHI)肉汤的情况下,将含有大肠杆菌 O157:H7 的样品循环通过系统 18 小时。大肠杆菌 O157:H7 的细胞特异性地捕获并富集在 QCM 的芯片表面,通过 QCM 频率变化来识别。李斯特菌和肠炎沙门氏菌被用作阴性对照。细菌富集后,添加检测抗体功能化的 AuNPs 以增强检测信号的变化。BHI 富集的使用进一步提高了开发系统的灵敏度,实现了 0-1 对数 CFU/ml 或 g 的检测限。本研究开发的实时监测活大肠杆菌 O157:H7 的方法可在 24 小时内同时富集和检测活菌。所开发系统的独特优势为常规食品样品中微生物分析提供了巨大的潜力。