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Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation

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Author(s): Michelle M. Packard | Elizabeth K. Wheeler | Evangelyn C. Alocilja | Maxim Shusteff

Journal: Diagnostics
ISSN 2075-4418

Volume: 3;
Issue: 1;
Start page: 105;
Date: 2013;
Original page

Keywords: thermal lysis | microfluidics | sample preparation | on-chip diagnostics | lab-on-a-chip | bacterial detection

ABSTRACT
Development of new diagnostic platforms that incorporate lab-on-a-chip technologies for portable assays is driving the need for rapid, simple, low cost methods to prepare samples for downstream processing or detection. An important component of the sample preparation process is cell lysis. In this work, a simple microfluidic thermal lysis device is used to quickly release intracellular nucleic acids and proteins without the need for additional reagents or beads used in traditional chemical or mechanical methods (e.g., chaotropic salts or bead beating). On-chip lysis is demonstrated in a multi-turn serpentine microchannel with external temperature control via an attached resistive heater. Lysis was confirmed for Escherichia coli by fluorescent viability assay, release of ATP measured with bioluminescent assay, release of DNA measured by fluorometry and qPCR, as well as bacterial culture. Results comparable to standard lysis techniques were achievable at temperatures greater than 65 °C and heating durations between 1 and 60 s.
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