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A disposable chemical heater and dry enzyme preparation for lysis and extraction of DNA and RNA from microorganisms
Sample preparation, including bacterial lysis, remains a hurdle in the realization of complete point-of-care tests for many pathogens. Here, we developed a sample preparation methodology for enzymatic lysis and sample heating for low-resource, point-of-care applications. We show an instrument-free c...
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| Published in: | Analytical methods 2016-04, Vol.8 (14), p.288-2886 |
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| Main Authors: | , , , , , , , , , , |
| Format: | Article |
| Language: | English |
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| cited_by | cdi_FETCH-LOGICAL-c448t-aedcf9fc50f4c66643a22ccf4de378e49f0846265c48807595d8cedb118844433 |
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| cites | cdi_FETCH-LOGICAL-c448t-aedcf9fc50f4c66643a22ccf4de378e49f0846265c48807595d8cedb118844433 |
| container_end_page | 2886 |
| container_issue | 14 |
| container_start_page | 288 |
| container_title | Analytical methods |
| container_volume | 8 |
| creator | Buser, J. R Zhang, X Byrnes, S. A Ladd, P. D Heiniger, E. K Wheeler, M. D Bishop, J. D Englund, J. A Lutz, B Weigl, B. H Yager, P |
| description | Sample preparation, including bacterial lysis, remains a hurdle in the realization of complete point-of-care tests for many pathogens. Here, we developed a sample preparation methodology for enzymatic lysis and sample heating for low-resource, point-of-care applications. We show an instrument-free chemical heater system for rapid lysis of a Gram-positive bacterium (
Staphylococcus aureus
) and an RNA virus (human respiratory syncytial virus) using a dried lysis enzyme mixture (achromopeptidase) for
S. aureus
. After a lysis step ( |
| doi_str_mv | 10.1039/c6ay00107f |
| format | article |
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Staphylococcus aureus
) and an RNA virus (human respiratory syncytial virus) using a dried lysis enzyme mixture (achromopeptidase) for
S. aureus
. After a lysis step (<1 minute), lysis enzymes are heat deactivated (<5 minutes) using a simple disposable chemical heater. We demonstrated that both DNA and RNA in the heat-treated sample could be directly amplified without purification, even in the presence of a clinically-obtained human nasal sample. This simple approach to dry enzyme storage and sample heating is adaptable to many applications where samples need to be lysed, including use in low-resource laboratories and in single-use or cartridge-based point-of-care diagnostic devices.
A simple dry enzyme and sample heating approach for point-of-care molecular diagnostic devices is demonstrated.</description><identifier>ISSN: 1759-9660</identifier><identifier>EISSN: 1759-9679</identifier><identifier>DOI: 10.1039/c6ay00107f</identifier><identifier>PMID: 37457919</identifier><language>eng</language><publisher>England</publisher><subject>Bacteria ; Drying ; Enzymes ; Heaters ; Heating ; Heating equipment ; Human respiratory syncytial virus ; Ribonucleic acids ; Staphylococcus aureus ; Viruses</subject><ispartof>Analytical methods, 2016-04, Vol.8 (14), p.288-2886</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c448t-aedcf9fc50f4c66643a22ccf4de378e49f0846265c48807595d8cedb118844433</citedby><cites>FETCH-LOGICAL-c448t-aedcf9fc50f4c66643a22ccf4de378e49f0846265c48807595d8cedb118844433</cites><orcidid>0000-0003-3555-8154</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/37457919$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Buser, J. R</creatorcontrib><creatorcontrib>Zhang, X</creatorcontrib><creatorcontrib>Byrnes, S. A</creatorcontrib><creatorcontrib>Ladd, P. D</creatorcontrib><creatorcontrib>Heiniger, E. K</creatorcontrib><creatorcontrib>Wheeler, M. D</creatorcontrib><creatorcontrib>Bishop, J. D</creatorcontrib><creatorcontrib>Englund, J. A</creatorcontrib><creatorcontrib>Lutz, B</creatorcontrib><creatorcontrib>Weigl, B. H</creatorcontrib><creatorcontrib>Yager, P</creatorcontrib><title>A disposable chemical heater and dry enzyme preparation for lysis and extraction of DNA and RNA from microorganisms</title><title>Analytical methods</title><addtitle>Anal Methods</addtitle><description>Sample preparation, including bacterial lysis, remains a hurdle in the realization of complete point-of-care tests for many pathogens. Here, we developed a sample preparation methodology for enzymatic lysis and sample heating for low-resource, point-of-care applications. We show an instrument-free chemical heater system for rapid lysis of a Gram-positive bacterium (
Staphylococcus aureus
) and an RNA virus (human respiratory syncytial virus) using a dried lysis enzyme mixture (achromopeptidase) for
S. aureus
. After a lysis step (<1 minute), lysis enzymes are heat deactivated (<5 minutes) using a simple disposable chemical heater. We demonstrated that both DNA and RNA in the heat-treated sample could be directly amplified without purification, even in the presence of a clinically-obtained human nasal sample. This simple approach to dry enzyme storage and sample heating is adaptable to many applications where samples need to be lysed, including use in low-resource laboratories and in single-use or cartridge-based point-of-care diagnostic devices.
A simple dry enzyme and sample heating approach for point-of-care molecular diagnostic devices is demonstrated.</description><subject>Bacteria</subject><subject>Drying</subject><subject>Enzymes</subject><subject>Heaters</subject><subject>Heating</subject><subject>Heating equipment</subject><subject>Human respiratory syncytial virus</subject><subject>Ribonucleic acids</subject><subject>Staphylococcus aureus</subject><subject>Viruses</subject><issn>1759-9660</issn><issn>1759-9679</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><recordid>eNqFkcFLwzAUxoMobk4v3pUcRZgmTZomxzGdCqIgevBUsvRFK21Tkw6sf71xm_O403u878cH3_sQOqbkghKmLo3QPSGUZHYHDWmWqrESmdrd7IIM0EEIH4QIxQTdRwOW8TRTVA1RmOCiDK0Lel4BNu9Ql0ZX-B10Bx7rpsCF7zE0330NuPXQaq-70jXYOo-rPpRhCcFX57VZCs7iq4fJ8voUp_WuxtHUO-ffdFOGOhyiPaurAEfrOUIvs-vn6e34_vHmbjq5HxvOZTfWUBirrEmJ5UYIwZlOEmMsL4BlEriyRHKRiNRwKUmMmhbSQDGnVErOOWMjdLbybb37XEDo8roMBqpKN-AWIU8kk9FVJNlWlEoi44MVV9vRTKYJJ5KlET1foTF9CB5s3vqy1r7PKcl_q8unYvK6rG4W4dO172JeQ7FB_7qKwMkK8MFs1P_u2Q-2mp15</recordid><startdate>20160414</startdate><enddate>20160414</enddate><creator>Buser, J. R</creator><creator>Zhang, X</creator><creator>Byrnes, S. A</creator><creator>Ladd, P. D</creator><creator>Heiniger, E. K</creator><creator>Wheeler, M. D</creator><creator>Bishop, J. D</creator><creator>Englund, J. A</creator><creator>Lutz, B</creator><creator>Weigl, B. H</creator><creator>Yager, P</creator><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>7SR</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>JG9</scope><scope>L7M</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0003-3555-8154</orcidid></search><sort><creationdate>20160414</creationdate><title>A disposable chemical heater and dry enzyme preparation for lysis and extraction of DNA and RNA from microorganisms</title><author>Buser, J. R ; Zhang, X ; Byrnes, S. A ; Ladd, P. D ; Heiniger, E. K ; Wheeler, M. D ; Bishop, J. D ; Englund, J. A ; Lutz, B ; Weigl, B. H ; Yager, P</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c448t-aedcf9fc50f4c66643a22ccf4de378e49f0846265c48807595d8cedb118844433</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Bacteria</topic><topic>Drying</topic><topic>Enzymes</topic><topic>Heaters</topic><topic>Heating</topic><topic>Heating equipment</topic><topic>Human respiratory syncytial virus</topic><topic>Ribonucleic acids</topic><topic>Staphylococcus aureus</topic><topic>Viruses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Buser, J. R</creatorcontrib><creatorcontrib>Zhang, X</creatorcontrib><creatorcontrib>Byrnes, S. A</creatorcontrib><creatorcontrib>Ladd, P. D</creatorcontrib><creatorcontrib>Heiniger, E. K</creatorcontrib><creatorcontrib>Wheeler, M. D</creatorcontrib><creatorcontrib>Bishop, J. D</creatorcontrib><creatorcontrib>Englund, J. A</creatorcontrib><creatorcontrib>Lutz, B</creatorcontrib><creatorcontrib>Weigl, B. H</creatorcontrib><creatorcontrib>Yager, P</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Materials Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Buser, J. R</au><au>Zhang, X</au><au>Byrnes, S. A</au><au>Ladd, P. D</au><au>Heiniger, E. K</au><au>Wheeler, M. D</au><au>Bishop, J. D</au><au>Englund, J. A</au><au>Lutz, B</au><au>Weigl, B. H</au><au>Yager, P</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A disposable chemical heater and dry enzyme preparation for lysis and extraction of DNA and RNA from microorganisms</atitle><jtitle>Analytical methods</jtitle><addtitle>Anal Methods</addtitle><date>2016-04-14</date><risdate>2016</risdate><volume>8</volume><issue>14</issue><spage>288</spage><epage>2886</epage><pages>288-2886</pages><issn>1759-9660</issn><eissn>1759-9679</eissn><abstract>Sample preparation, including bacterial lysis, remains a hurdle in the realization of complete point-of-care tests for many pathogens. Here, we developed a sample preparation methodology for enzymatic lysis and sample heating for low-resource, point-of-care applications. We show an instrument-free chemical heater system for rapid lysis of a Gram-positive bacterium (
Staphylococcus aureus
) and an RNA virus (human respiratory syncytial virus) using a dried lysis enzyme mixture (achromopeptidase) for
S. aureus
. After a lysis step (<1 minute), lysis enzymes are heat deactivated (<5 minutes) using a simple disposable chemical heater. We demonstrated that both DNA and RNA in the heat-treated sample could be directly amplified without purification, even in the presence of a clinically-obtained human nasal sample. This simple approach to dry enzyme storage and sample heating is adaptable to many applications where samples need to be lysed, including use in low-resource laboratories and in single-use or cartridge-based point-of-care diagnostic devices.
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| ispartof | Analytical methods, 2016-04, Vol.8 (14), p.288-2886 |
| issn | 1759-9660 1759-9679 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_1785240835 |
| source | Royal Society of Chemistry |
| subjects | Bacteria Drying Enzymes Heaters Heating Heating equipment Human respiratory syncytial virus Ribonucleic acids Staphylococcus aureus Viruses |
| title | A disposable chemical heater and dry enzyme preparation for lysis and extraction of DNA and RNA from microorganisms |
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