Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Packard, Michelle M., Wheeler, Elizabeth K., Alocilja, Evangelyn C., Shusteff, Maxim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2013
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
_version_ 1782403600478109696
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
work_keys_str_mv AT packardmichellem performanceevaluationoffastmicrofluidicthermallysisofbacteriafordiagnosticsamplepreparation
AT wheelerelizabethk performanceevaluationoffastmicrofluidicthermallysisofbacteriafordiagnosticsamplepreparation
AT alociljaevangelync performanceevaluationoffastmicrofluidicthermallysisofbacteriafordiagnosticsamplepreparation
AT shusteffmaxim performanceevaluationoffastmicrofluidicthermallysisofbacteriafordiagnosticsamplepreparation