Abstract
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.
Highlights
During water electrolysis, the micro-environment at the electrode/electrolyte interface has different properties compare to that of the bulk electrolyte
10 min and 15 min of electrochemical cell lysis (ECL), respectively. These results show that the bacteria in both pond water and wastewater were rapidly and efficiently lysed by ECL with DCT values comparable to those obtained with the commercial kit
The successful application of ECL on different environmental water samples suggests the potential application of ECL as a rapid and reagent-free sample preparation technique with a low voltage requirement for microbial monitoring in the field
Summary
The micro-environment at the electrode/electrolyte interface has different properties compare to that of the bulk electrolyte. Lower electroporation voltages can be realized using nano-structured electrodes coupled with microfluidic devices This approach would require a complicated fabrication process and precise operation [11,13e15,17,23e25]. We note that the aforementioned studies of ECL were mainly focused on clinical samples (e.g., human cells [7,26]), and conducted in well-controlled systems with purified buffers. All of these studies highlighted in the development of micro-scale devices with microliter or even nanoliter throughput. Our overarching goal is to determine the DNA extraction efficiencies as a function of the key operational parameters (i.e., pH ranges with varied treatment durations) for the use of ECL, as applied to DNA extraction and PCR amplification of gram-positive and gram-negative bacteria in real surface water and wastewater
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