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Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation†
Nano-Biosensors Laboratory, Michigan State University, 115 Farrall Hall, East Lansing, MI 48824, USA
Lawrence Livermore National Laboratory, 7000 East Ave, Livermore, CA 94450, USA
This author deceased on 4 July 2012.
† This work was performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344, LLNL-JRNL-586013.
* Author to whom correspondence should be addressed.
Received: 10 December 2012; in revised form: 10 January 2013 / Accepted: 16 January 2013 / Published: 17 January 2013
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.
Keywords: thermal lysis; microfluidics; sample preparation; on-chip diagnostics; lab-on-a-chip; bacterial detection
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Packard, M.M.; Wheeler, E.K.; Alocilja, E.C.; Shusteff, M. Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation. Diagnostics 2013, 3, 105-116.
Packard MM, Wheeler EK, Alocilja EC, Shusteff M. Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation. Diagnostics. 2013; 3(1):105-116.
Packard, Michelle M.; Wheeler, Elizabeth K.; Alocilja, Evangelyn C.; Shusteff, Maxim. 2013. "Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation." Diagnostics 3, no. 1: 105-116.