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Electrochemical cell lysis of gram-positive and gram-negative bacteria: DNA extraction from environmental water samples

Wang, Siwen and Zhu, Yanzhe and Yang, Yang and Li, Jing and Hoffmann, Michael R. (2020) Electrochemical cell lysis of gram-positive and gram-negative bacteria: DNA extraction from environmental water samples. Electrochimica Acta, 338 . Art. No. 135864. ISSN 0013-4686. PMCID PMC7063685. doi:10.1016/j.electacta.2020.135864.

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Cell lysis is an essential step for the nucleic acid-based surveillance of bacteriological water quality. Recently, electrochemical cell lysis (ECL), which is based on the local generation of hydroxide at a cathode surface, has been reported to be a rapid and reagent-free method for cell lysis. Herein, we describe the development of a milliliter-output ECL device and its performance characterization with respect to the DNA extraction efficiency for gram-negative bacteria (Escherichia coli and Salmonella Typhi) and gram-positive bacteria (Enterococcus durans and Bacillus subtilis). Both gram-negative and gram-positive bacteria were successfully lysed within a short but optimal duration of 1 min at a low voltage of ∼5 V. The ECL method described herein, is demonstrated to be applicable to various environmental water sample types, including pond water, treated wastewater, and untreated wastewater with DNA extraction efficiencies similar to a commercial DNA extraction kit. The ECL system outperformed homogeneous chemical lysis in terms of reaction times and DNA extraction efficiencies, due in part to the high pH generated at the cathode surface, which was predicted by simulations of the hydroxide transport in the cathodic chamber. Our work indicates that the ECL method for DNA extraction is rapid, simplified and low-cost with no need for complex instrumentation. It has demonstrable potential as a prelude to PCR analyses of waterborne bacteria in the field, especially for the gram-negative ones.

Item Type:Article
Related URLs:
URLURL TypeDescription CentralArticle
Wang, Siwen0000-0002-8553-425X
Zhu, Yanzhe0000-0002-2260-1830
Yang, Yang0000-0003-3767-8029
Li, Jing0000-0003-0639-9422
Hoffmann, Michael R.0000-0002-0432-6564
Additional Information:© 2020 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license ( Received 28 October 2019, Revised 4 February 2020, Accepted 5 February 2020, Available online 6 February 2020. This research was supported by the Bill and Melinda Gates Foundation, Seattle, WA [OPP1111252]. The authors acknowledge Xingyu Lin, Katharina Urmann and Jared R. Leadbetter, for their helpful discussion and critical reviews of our manuscript. We also thank Clement Cid and Nissim Gore-Datar for their help on the sampling of latrine wastewater. CRediT authorship contribution statement: Yanzhe Zhu: Writing - original draft, Writing - review & editing. Yang Yang: Writing - review & editing. Jing Li: Formal analysis, Writing - review & editing. Michael R. Hoffmann: Writing - review & editing. There are no conflicts of interest to declare.
Funding AgencyGrant Number
Bill and Melinda Gates FoundationOPP1111252
Subject Keywords:Electrochemical cell lysis (ECL); DNA extraction; Gram-positive and gram-negative bacteria; Environmental water samples
PubMed Central ID:PMC7063685
Record Number:CaltechAUTHORS:20200206-125056339
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Official Citation:Siwen Wang, Yanzhe Zhu, Yang Yang, Jing Li, Michael R. Hoffmann, Electrochemical cell lysis of gram-positive and gram-negative bacteria: DNA extraction from environmental water samples, Electrochimica Acta, Volume 338, 2020, 135864, ISSN 0013-4686,
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:101162
Deposited By: George Porter
Deposited On:07 Feb 2020 19:09
Last Modified:15 Feb 2022 23:42

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