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Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation (†)
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 w...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4665579/ https://www.ncbi.nlm.nih.gov/pubmed/26835670 http://dx.doi.org/10.3390/diagnostics3010105 |
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author | Packard, Michelle M. Wheeler, Elizabeth K. Alocilja, Evangelyn C. Shusteff, Maxim |
author_facet | Packard, Michelle M. Wheeler, Elizabeth K. Alocilja, Evangelyn C. Shusteff, Maxim |
author_sort | Packard, Michelle M. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4665579 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-46655792016-01-27 Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation (†) Packard, Michelle M. Wheeler, Elizabeth K. Alocilja, Evangelyn C. Shusteff, Maxim Diagnostics (Basel) Communication 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. MDPI 2013-01-17 /pmc/articles/PMC4665579/ /pubmed/26835670 http://dx.doi.org/10.3390/diagnostics3010105 Text en © 2013 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/). |
spellingShingle | Communication Packard, Michelle M. Wheeler, Elizabeth K. Alocilja, Evangelyn C. Shusteff, Maxim Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation (†) |
title | Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation (†) |
title_full | Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation (†) |
title_fullStr | Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation (†) |
title_full_unstemmed | Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation (†) |
title_short | Performance Evaluation of Fast Microfluidic Thermal Lysis of Bacteria for Diagnostic Sample Preparation (†) |
title_sort | performance evaluation of fast microfluidic thermal lysis of bacteria for diagnostic sample preparation (†) |
topic | Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4665579/ https://www.ncbi.nlm.nih.gov/pubmed/26835670 http://dx.doi.org/10.3390/diagnostics3010105 |
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