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基于自供电微球传感器的微生物及其对抗生素响应的无标记监测。

Label-Free Monitoring of Microorganisms and Their Responses to Antibiotics Based on Self-Powered Microbead Sensors.

机构信息

Division of Urology, Department of Surgery , Chi Mei Medical Center , Tainan City , Taiwan 710.

Department of Orthopedic Surgery, National Cheng Kung University Hospital, College of Medicine , National Cheng Kung University , Tainan City , Taiwan 701.

出版信息

ACS Sens. 2018 Oct 26;3(10):2182-2190. doi: 10.1021/acssensors.8b00790. Epub 2018 Oct 1.

Abstract

Rapid detection of bacteria and their susceptibility to specific antibiotics plays a vital role in microbial infection treatments. Antimicrobial susceptibility testing (AST) is a common measure to select effective drugs. However, the conventional practices, such as broth dilution, E-test, and disk diffusion, in clinical applications require a long turnaround time (∼3 days), thereby compromising treatments and increasing mortality. This study presents self-powered sensors for on-site microorganism monitoring and rapid AST based on functionalized microbeads. The microbead sensors are driven by Brownian motion, rendering external power unnecessary. Fluorescent microbeads ( d = 2 μm) were coated with vancomycin to capture bacteria. The growth and responses of Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus under antibiotic treatment were evaluated. The method showed stable selective binding despite the presence of some interferential substances, such as proteins and cells. Diffusivity change was strongly related to bacterial concentration. Accordingly, the diffusivity values of microbeads bound with motile and nonmotile bacteria exhibited specific patterns because of extra motility from microbes and increased particle diameter. Only a drop of microbead-bacteria suspension (∼5 μL) was needed in a microchip for each measurement. The microchip provided a steady environment for measurement over a few hours. By distinguishing the slope of the last four data points in the temporal diffusivity curve, bacterial susceptibility or resistance to specific antibiotics could be determined within a time frame of 2 h. The study provides insights into saving more lives by using a fast and robust AST technique in future clinical practice.

摘要

快速检测细菌及其对特定抗生素的敏感性在微生物感染治疗中起着至关重要的作用。抗菌药物敏感性测试(AST)是选择有效药物的常用方法。然而,临床应用中的常规方法,如肉汤稀释法、E 试验和药敏纸片扩散法,需要较长的周转时间(~3 天),从而影响治疗效果并增加死亡率。本研究提出了基于功能化微球的现场微生物监测和快速 AST 的自供电传感器。微球传感器由布朗运动驱动,无需外部电源。荧光微球(d=2μm)用万古霉素包被以捕获细菌。评估了抗生素处理下革兰氏阴性大肠杆菌和革兰氏阳性金黄色葡萄球菌的生长和反应。该方法表现出稳定的选择性结合,尽管存在一些干扰物质,如蛋白质和细胞。扩散率的变化与细菌浓度密切相关。因此,由于微生物的额外运动和颗粒直径的增加,与运动和非运动细菌结合的微球的扩散率值表现出特定的模式。每次测量只需要微芯片中约 5μL 的微球-细菌悬浮液。微芯片为几个小时的测量提供了稳定的环境。通过区分时间扩散率曲线上最后四个数据点的斜率,可以在 2 小时内确定细菌对特定抗生素的敏感性或耐药性。该研究为未来临床实践中使用快速、稳健的 AST 技术挽救更多生命提供了思路。

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