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基于原子层沉积(ALD)氧化铝钝化的微电极,通过裂解酶作为选择性手段,实现了对全细胞细菌的无标记、微流控和阻抗检测。

Lytic enzymes as selectivity means for label-free, microfluidic and impedimetric detection of whole-cell bacteria using ALD-Al2O3 passivated microelectrodes.

机构信息

ICTEAM Institute, Université catholique de Louvain (UCL), Place du levant 3-L5.03.02, 1348 Louvain-la-Neuve, Belgium.

ELI Institute Applied Microbiology (ELIM), Université catholique de Louvain (UCL), Croix du Sud 2-L7.05.12, 1348 Louvain-la-Neuve, Belgium.

出版信息

Biosens Bioelectron. 2015 May 15;67:154-61. doi: 10.1016/j.bios.2014.07.084. Epub 2014 Aug 7.

DOI:10.1016/j.bios.2014.07.084
PMID:25149092
Abstract

Point-of-care (PoC) diagnostics for bacterial detection offer tremendous prospects for public health care improvement. However, such tools require the complex combination of the following performances: rapidity, selectivity, sensitivity, miniaturization and affordability. To meet these specifications, this paper presents a new selectivity method involving lysostaphin together with a CMOS-compatible impedance sensor for genus-specific bacterial detection. The method enables the sample matrix to be directly flown on the polydopamine-covered sensor surface without any pre-treatment, and considerably reduces the background noise. Experimental proof-of-concept, explored by simulations and confirmed through a setup combining simultaneous optical and electrical real-time monitoring, illustrates the selective and capacitive detection of Staphylococcus epidermidis in synthetic urine also containing Enterococcus faecium. While providing capabilities for miniaturization and system integration thanks to CMOS compatibility, the sensors show a detection limit of ca. 10(8) (CFU/mL).min in a 1.5 μL microfluidic chamber with an additional setup time of 50 min. The potentials, advantages and limitations of the method are also discussed.

摘要

即时检测(PoC)诊断技术在细菌检测方面具有巨大的应用前景,能够极大地改善公共医疗保健水平。然而,此类工具需要以下性能的复杂结合:快速性、选择性、灵敏度、小型化和可负担性。为了满足这些要求,本文提出了一种新的选择性方法,该方法涉及溶菌酶与 CMOS 兼容的阻抗传感器结合,用于属特异性细菌检测。该方法可使样本基质无需任何预处理即可直接在聚多巴胺覆盖的传感器表面流动,并显著降低背景噪声。通过仿真实验和通过结合光学和电气实时监测的设置进行的实验验证,说明了在含有粪肠球菌的合成尿液中对表皮葡萄球菌的选择性和电容检测。该传感器由于与 CMOS 的兼容性而具有小型化和系统集成的能力,在具有 50 分钟额外设置时间的 1.5 μL 微流控室中,检测限约为 10(8)(CFU/mL).min。本文还讨论了该方法的潜力、优势和局限性。

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