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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2018
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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 |
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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 |
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