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介电泳辅助 G-FETs 快速、选择性和单细胞检测抗生素耐药菌。

Dielectrophoresis assisted rapid, selective and single cell detection of antibiotic resistant bacteria with G-FETs.

机构信息

Department of Physics, Boston College, Chestnut Hill, MA, 02467, United States.

Department of Chemistry, Boston College, Chestnut Hill, MA, 02467, United States.

出版信息

Biosens Bioelectron. 2020 May 15;156:112123. doi: 10.1016/j.bios.2020.112123. Epub 2020 Feb 27.

Abstract

The rapid increase in antibiotic resistant pathogenic bacteria has become a global threat, which besides the development of new drugs, requires rapid, cheap, scalable, and accurate diagnostics. Label free biosensors relying on electrochemical, mechanical, and mass based detection of whole bacterial cells have attempted to meet these requirements. However, the trade-off between selectivity and sensitivity of such sensors remains a key challenge. In particular, point-of-care diagnostics that are able to reduce and/or prevent unneeded antibiotic prescriptions require highly specific probes with sensitive and accurate transducers that can be miniaturized and multiplexed, and that are easy to operate and cheap. Towards achieving this goal, we present a number of advances in the use of graphene field effect transistors (G-FET) including the first use of peptide probes to electrically detect antibiotic resistant bacteria in a highly specific manner. In addition, we dramatically reduce the needed concentration for detection by employing dielectrophoresis for the first time in a G-FET, allowing us to monitor changes in the Dirac point due to individual bacterial cells. Specifically, we realized rapid binding of bacterial cells to a G-FET by electrical field guiding to the device to realize an overall 3 orders of magnitude decrease in cell-concentration enabling a single-cell detection limit, and 9-fold reduction in needed time to 5 min. Utilizing our new biosensor and procedures, we demonstrate the first selective, electrical detection of the pathogenic bacterial species Staphylococcus aureus and antibiotic resistant Acinetobacter baumannii on a single platform.

摘要

抗生素耐药病原菌的迅速增加已成为全球性威胁,除了开发新药物外,还需要快速、廉价、可扩展和准确的诊断方法。依赖于电化学、机械和基于质量的全细菌细胞检测的无标记生物传感器已经尝试满足这些要求。然而,此类传感器的选择性和灵敏度之间的权衡仍然是一个关键挑战。特别是,能够减少和/或预防不必要的抗生素处方的即时诊断需要具有高特异性的探针,这些探针具有敏感和准确的换能器,能够小型化和多重化,并且易于操作且廉价。为了实现这一目标,我们在使用石墨烯场效应晶体管(G-FET)方面取得了一些进展,包括首次使用肽探针以高度特异性的方式电检测抗生素耐药细菌。此外,我们通过首次在 G-FET 中使用介电泳显著降低了检测所需的浓度,从而能够监测由于单个细菌细胞而引起的狄拉克点的变化。具体来说,我们通过电场引导到器件上来实现细菌细胞与 G-FET 的快速结合,从而实现细胞浓度降低 3 个数量级,实现单细胞检测极限,并将所需时间缩短 9 倍,达到 5 分钟。利用我们的新生物传感器和程序,我们在单个平台上首次选择性地、电检测到致病细菌物种金黄色葡萄球菌和抗生素耐药鲍曼不动杆菌。

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