Department of Clinical Chemistry, Faculty of Medical Technology, Mahidol University, Nakhon Pathom 73170, Thailand.
Biosens Bioelectron. 2010 Oct 15;26(2):784-9. doi: 10.1016/j.bios.2010.06.024. Epub 2010 Jul 1.
This work presents the first demonstration of a cantilever based cholerae sensor. Dynamic force microscopy within atomic force microscope (AFM) is applied to measure the cantilever's resonance frequency shift due to mass of cell bound on microcantilever surface. The Vibrio cholerae O1, a food and waterborne pathogen that caused cholera disease in human, is a target bacterium cell of interest. Commercial gold-coated AFM microcantilevers are immobilized with monoclonal antibody (anti-V. cholerae O1) by self-assembled monolayer method. V. cholerae O1 detection experiment is then conducted in concentrations ranging from 1×10(3) to 1×10(7) CFU/ml. The microcantilever-based sensor has a detection limit of ∼1×10(3) CFU/ml and a mass sensitivity, Δm/ΔF, of ∼146.5 pg/Hz, which is at least two orders of magnitude lower than other reported techniques and sufficient for V. cholerae detection in food products without pre-enrichment steps. In addition, V. cholerae O1 antigen-antibody binding on microcanilever is confirmed by scanning electron microscopy. The results demonstrate that the new biosensor is promising for high sensitivity, uncomplicated and rapid detection of V. cholerae O1.
本工作首次展示了基于悬臂梁的霍乱传感器。原子力显微镜(AFM)中的动态力显微镜用于测量由于微悬臂表面上结合的细胞质量而导致的悬臂共振频率的偏移。霍乱弧菌 O1 是一种食源性和水源性病原体,可引起人类霍乱病,是感兴趣的目标细菌细胞。商用金涂层 AFM 微悬臂通过自组装单层方法固定单克隆抗体(抗霍乱弧菌 O1)。然后在 1×10(3)至 1×10(7)CFU/ml 的浓度范围内进行霍乱弧菌 O1 检测实验。基于微悬臂的传感器的检测限约为 1×10(3)CFU/ml,质量灵敏度Δm/ΔF 约为 146.5 pg/Hz,这至少比其他报道的技术低两个数量级,足以在没有预富集步骤的情况下检测食品中的霍乱弧菌 O1。此外,通过扫描电子显微镜证实了微悬臂上霍乱弧菌 O1 抗原-抗体的结合。结果表明,新型生物传感器有望实现对霍乱弧菌 O1 的高灵敏度、简单快速的检测。