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Title: Single Escherichia coli bacteria detection using a chemiluminescence digital microwell array chip
Authors: Wu, Wenshuai
Nguyen, Binh Thi Thanh
Liu, Patricia Yang
Cai, Gaozhe
Feng, Shilun
Shi, Yuzhi
Zhang, Boran
Hong, Yuzhi
Yu, Ruozhen
Zhou, Xiaohong
Liu, Ai Qun
Zhang, Yi
Yap, Eric Peng Huat
Chin, Lip Ket
Keywords: Engineering::Electrical and electronic engineering
Issue Date: 2022
Source: Wu, W., Nguyen, B. T. T., Liu, P. Y., Cai, G., Feng, S., Shi, Y., Zhang, B., Hong, Y., Yu, R., Zhou, X., Liu, A. Q., Zhang, Y., Yap, E. P. H. & Chin, L. K. (2022). Single Escherichia coli bacteria detection using a chemiluminescence digital microwell array chip. Biosensors & Bioelectronics, 215, 114594-.
Project: MOE2017-T3-1-001
Journal: Biosensors & Bioelectronics
Abstract: Rapid and sensitive Escherichia coli (E. coli) detection is important in determining environmental contamination, food contamination, as well as bacterial infection. Conventional methods based on bacterial culture suffer from long testing time (24 h), whereas novel nucleic acid-based and immunolabelling approaches are hindered by complicated operation, the need of complex and costly equipment, and the lack of differentiation of live and dead bacteria. Herein, we propose a chemiluminescence digital microwell array chip based on the hydrolysis of 6-Chloro-4-methylumbelliferyl-β-D-glucuronide by the β-D-glucuronidase in E. coli to achieve fast single bacterial fluorescence detection. Taking the advantage of the picoliter microwells, single bacteria are digitally encapsulated in these microwells, thus the accurate quantification of E. coli can be realized by counting the number of positive microwells. We also show that the chemiluminescence digital microwell array chip is not affected by the turbidity of the test samples as well as the temperature. Most importantly, our method can differentiate live and dead bacteria through bacterial proliferation and enzyme expression, which is confirmed by detecting E. coli after pH and chlorination treatment. By comparing with the standard method of plate counting, our method has comparable performance but significantly reduces the testing time from over 24 h-2 h and 4 h for qualitative and quantitative analysis, respectively. In addition, the microfluidic chip is portable and easy to operate without external pump, which is promising as a rapid and on-site platform for single E. coli analysis in water and food monitoring, as well as infection diagnosis.
ISSN: 2155-6210
DOI: 10.1016/j.bios.2022.114594
Rights: © 2022 Elsevier B.V. All rights reserved.
Fulltext Permission: none
Fulltext Availability: No Fulltext
Appears in Collections:EEE Journal Articles
LKCMedicine Journal Articles

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