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 conventional...
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| Veröffentlicht in: | PloS one 2018-07, Vol.13 (7), p.e0201070-e0201070 |
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| creator | Santaus, Tonya M Li, Shan Ladd, Paula Harvey, Amanda Cole, Shannon Stine, O Colin Geddes, Chris D |
| 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 |
| doi_str_mv | 10.1371/journal.pone.0201070 |
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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.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0201070</identifier><identifier>PMID: 30044836</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Animals ; Bacteria ; Bacteriological Techniques ; Biochemistry ; Biodegradation ; Biology and Life Sciences ; Chlamydia ; Degradation ; Deoxyribonucleic acid ; Digital Object Identifier ; DNA ; DNA fragmentation ; DNA, Bacterial ; Drug therapy ; Epidemiology ; Gold ; Intracellular ; Irradiation ; Listeria ; Listeria monocytogenes ; Listeria monocytogenes - genetics ; Listeria monocytogenes - isolation & purification ; Lysis ; Medical research ; Medicine and Health Sciences ; Microwaves ; Physical Sciences ; Polymerase Chain Reaction ; Proteins ; Public health ; Research and Analysis Methods ; Salmonella ; Sample preparation ; Sheep ; Temperature ; Viability ; Vibrio cholerae ; Vibrio cholerae - genetics ; Vibrio cholerae - isolation & purification ; Water-borne diseases ; Waterborne diseases</subject><ispartof>PloS one, 2018-07, Vol.13 (7), p.e0201070-e0201070</ispartof><rights>COPYRIGHT 2018 Public Library of Science</rights><rights>2018 Santaus et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2018 Santaus et al 2018 Santaus et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-8774fd429e6bd0e3d80a0cbbef658fc1baaa837fb9fc9be8a258423f2d5b8de33</citedby><cites>FETCH-LOGICAL-c692t-8774fd429e6bd0e3d80a0cbbef658fc1baaa837fb9fc9be8a258423f2d5b8de33</cites><orcidid>0000-0002-3200-0393 ; 0000-0003-1805-1407 ; 0000-0003-3974-3752 ; 0000-0002-8643-2343</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6059484/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6059484/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793,79600,79601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30044836$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Mishra, Yogendra Kumar</contributor><creatorcontrib>Santaus, Tonya M</creatorcontrib><creatorcontrib>Li, Shan</creatorcontrib><creatorcontrib>Ladd, Paula</creatorcontrib><creatorcontrib>Harvey, Amanda</creatorcontrib><creatorcontrib>Cole, Shannon</creatorcontrib><creatorcontrib>Stine, O Colin</creatorcontrib><creatorcontrib>Geddes, Chris D</creatorcontrib><title>Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae</title><title>PloS one</title><addtitle>PLoS One</addtitle><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.</description><subject>Animals</subject><subject>Bacteria</subject><subject>Bacteriological Techniques</subject><subject>Biochemistry</subject><subject>Biodegradation</subject><subject>Biology and Life Sciences</subject><subject>Chlamydia</subject><subject>Degradation</subject><subject>Deoxyribonucleic acid</subject><subject>Digital Object Identifier</subject><subject>DNA</subject><subject>DNA fragmentation</subject><subject>DNA, Bacterial</subject><subject>Drug therapy</subject><subject>Epidemiology</subject><subject>Gold</subject><subject>Intracellular</subject><subject>Irradiation</subject><subject>Listeria</subject><subject>Listeria monocytogenes</subject><subject>Listeria monocytogenes - genetics</subject><subject>Listeria monocytogenes - isolation & purification</subject><subject>Lysis</subject><subject>Medical research</subject><subject>Medicine and Health Sciences</subject><subject>Microwaves</subject><subject>Physical Sciences</subject><subject>Polymerase Chain Reaction</subject><subject>Proteins</subject><subject>Public health</subject><subject>Research and Analysis Methods</subject><subject>Salmonella</subject><subject>Sample preparation</subject><subject>Sheep</subject><subject>Temperature</subject><subject>Viability</subject><subject>Vibrio cholerae</subject><subject>Vibrio cholerae - 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| source | Public Library of Science (PLoS) Journals Open Access; MEDLINE; DOAJ Directory of Open Access Journals; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Animals Bacteria Bacteriological Techniques Biochemistry Biodegradation Biology and Life Sciences Chlamydia Degradation Deoxyribonucleic acid Digital Object Identifier DNA DNA fragmentation DNA, Bacterial Drug therapy Epidemiology Gold Intracellular Irradiation Listeria Listeria monocytogenes Listeria monocytogenes - genetics Listeria monocytogenes - isolation & purification Lysis Medical research Medicine and Health Sciences Microwaves Physical Sciences Polymerase Chain Reaction Proteins Public health Research and Analysis Methods Salmonella Sample preparation Sheep Temperature Viability Vibrio cholerae Vibrio cholerae - genetics Vibrio cholerae - isolation & purification Water-borne diseases Waterborne diseases |
| title | Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae |
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