Cargando…

Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae

Sample preparation is a leading bottleneck in rapid detection of pathogenic bacteria. Here, we use Lyse-It(®) for bacterial cellular lysis, genomic DNA fragmentation, and protein release and degradation for both Listeria monocytogenes and Vibrio cholerae. The concept of Lyse-It(®) employs a conventi...

Descripción completa

Detalles Bibliográficos
Autores principales: Santaus, Tonya M., Li, Shan, Ladd, Paula, Harvey, Amanda, Cole, Shannon, Stine, O. Colin, Geddes, Chris D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6059484/
https://www.ncbi.nlm.nih.gov/pubmed/30044836
http://dx.doi.org/10.1371/journal.pone.0201070
_version_ 1783341871895937024
author Santaus, Tonya M.
Li, Shan
Ladd, Paula
Harvey, Amanda
Cole, Shannon
Stine, O. Colin
Geddes, Chris D.
author_facet Santaus, Tonya M.
Li, Shan
Ladd, Paula
Harvey, Amanda
Cole, Shannon
Stine, O. Colin
Geddes, Chris D.
author_sort Santaus, Tonya M.
collection PubMed
description Sample preparation is a leading bottleneck in rapid detection of pathogenic bacteria. Here, we use Lyse-It(®) for bacterial cellular lysis, genomic DNA fragmentation, and protein release and degradation for both Listeria monocytogenes and Vibrio cholerae. The concept of Lyse-It(®) employs a conventional microwave and Lyse-It(®) slides for intensely focused microwave irradiation onto the sample. High microwave power and a <60 second irradiation time allow for rapid cellular lysis and subsequent intracellular component release. The pathogenic bacteria are identified by quantitative polymerase chain reaction (qPCR), which subsequently demonstrates the viability of DNA for amplification post microwave-induced lysis. Intracellular component release, degradation, and detection of L. monocytogenes and V. cholerae has been performed and shown in this paper. These results demonstrate a rapid, low-cost, and efficient way for bacterial sample preparation on both food and water-borne Gram-positive and -negative organisms alike.
format Online
Article
Text
id pubmed-6059484
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-60594842018-08-09 Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae Santaus, Tonya M. Li, Shan Ladd, Paula Harvey, Amanda Cole, Shannon Stine, O. Colin Geddes, Chris D. PLoS One Research Article Sample preparation is a leading bottleneck in rapid detection of pathogenic bacteria. Here, we use Lyse-It(®) for bacterial cellular lysis, genomic DNA fragmentation, and protein release and degradation for both Listeria monocytogenes and Vibrio cholerae. The concept of Lyse-It(®) employs a conventional microwave and Lyse-It(®) slides for intensely focused microwave irradiation onto the sample. High microwave power and a <60 second irradiation time allow for rapid cellular lysis and subsequent intracellular component release. The pathogenic bacteria are identified by quantitative polymerase chain reaction (qPCR), which subsequently demonstrates the viability of DNA for amplification post microwave-induced lysis. Intracellular component release, degradation, and detection of L. monocytogenes and V. cholerae has been performed and shown in this paper. These results demonstrate a rapid, low-cost, and efficient way for bacterial sample preparation on both food and water-borne Gram-positive and -negative organisms alike. Public Library of Science 2018-07-25 /pmc/articles/PMC6059484/ /pubmed/30044836 http://dx.doi.org/10.1371/journal.pone.0201070 Text en © 2018 Santaus et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Santaus, Tonya M.
Li, Shan
Ladd, Paula
Harvey, Amanda
Cole, Shannon
Stine, O. Colin
Geddes, Chris D.
Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae
title Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae
title_full Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae
title_fullStr Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae
title_full_unstemmed Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae
title_short Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae
title_sort rapid sample preparation with lyse-it® for listeria monocytogenes and vibrio cholerae
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6059484/
https://www.ncbi.nlm.nih.gov/pubmed/30044836
http://dx.doi.org/10.1371/journal.pone.0201070
work_keys_str_mv AT santaustonyam rapidsamplepreparationwithlyseitforlisteriamonocytogenesandvibriocholerae
AT lishan rapidsamplepreparationwithlyseitforlisteriamonocytogenesandvibriocholerae
AT laddpaula rapidsamplepreparationwithlyseitforlisteriamonocytogenesandvibriocholerae
AT harveyamanda rapidsamplepreparationwithlyseitforlisteriamonocytogenesandvibriocholerae
AT coleshannon rapidsamplepreparationwithlyseitforlisteriamonocytogenesandvibriocholerae
AT stineocolin rapidsamplepreparationwithlyseitforlisteriamonocytogenesandvibriocholerae
AT geddeschrisd rapidsamplepreparationwithlyseitforlisteriamonocytogenesandvibriocholerae