Interdisciplinary Microsystems Group, Department of Mechanical and Aerospace Engineering, University of Florida, P.O. Box 116250, Gainesville, FL, 32611, USA.
Department of Environmental Engineering Sciences, University of Florida, P.O. Box 116580, Gainesville, FL, 32611, USA.
Sci Rep. 2023 Mar 14;13(1):4245. doi: 10.1038/s41598-023-31208-4.
Escherichia coli (E. coli) cells are present in fecal materials that can be the main source for disease-causing agents in water. As a result, E. coli is recommended as a water quality indicator. We have developed an innovative platform to detect E. coli for monitoring water quality on-site by integrating paper-based sample preparation with nucleic acid isothermal amplification. The platform carries out bacterial lysis and DNA enrichment onto a paper pad through ball-based valves for fluid control, with no need of laboratory equipment, followed by loop-mediated isothermal amplification (LAMP) in a battery-operated coffee mug, and colorimetric detection. We have used the platform to detect E. coli in environmental water samples in about 1 h, with a limit of quantitation of 0.2 CFU/mL, and 3 copies per reaction. The platform was confirmed for detecting multiple E. coli strains, and for water samples of different salt concentrations. We validated the functions of the platform by analyzing recreational water samples collected near the Atlantic Ocean that contain different concentrations of salt and bacteria.
大肠杆菌(E. coli)细胞存在于粪便物质中,可能是水中致病因子的主要来源。因此,大肠杆菌被推荐作为水质指标。我们开发了一种创新的平台,通过将基于纸张的样品制备与核酸等温扩增相结合,用于现场水质监测。该平台通过基于球的阀进行细菌裂解和 DNA 富集到纸垫上,用于流体控制,无需实验室设备,然后在电池供电的咖啡杯内进行环介导等温扩增(LAMP),并进行比色检测。我们已经使用该平台在大约 1 小时内检测到环境水样中的大肠杆菌,定量下限为 0.2 CFU/mL,每个反应 3 个拷贝。该平台已被证实可用于检测多种大肠杆菌菌株和不同盐浓度的水样。我们通过分析大西洋附近采集的娱乐用水样来验证该平台的功能,这些水样含有不同浓度的盐和细菌。