A bench-top automated workstation for nucleic acid isolation from clinical sample types
Systems that automate extraction of nucleic acid from cells or viruses in complex clinical matrices have tremendous value even in the absence of an integrated downstream detector. We describe our bench-top automated workstation that integrates our previously-reported extraction method – TruTip – wit...
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Veröffentlicht in: | Journal of microbiological methods 2018-05, Vol.148, p.174-180 |
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container_title | Journal of microbiological methods |
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creator | Thakore, Nitu Garber, Steve Bueno, Arial Qu, Peter Norville, Ryan Villanueva, Michael Chandler, Darrell P. Holmberg, Rebecca Cooney, Christopher G. |
description | Systems that automate extraction of nucleic acid from cells or viruses in complex clinical matrices have tremendous value even in the absence of an integrated downstream detector. We describe our bench-top automated workstation that integrates our previously-reported extraction method – TruTip – with our newly-developed mechanical lysis method. This is the first report of this method for homogenizing viscous and heterogeneous samples and lysing difficult-to-disrupt cells using "MagVor": a rotating magnet that rotates a miniature stir disk amidst glass beads confined inside of a disposable tube. Using this system, we demonstrate automated nucleic acid extraction from methicillin-resistant Staphylococcus aureus (MRSA) in nasopharyngeal aspirate (NPA), influenza A in nasopharyngeal swabs (NPS), human genomic DNA from whole blood, and Mycobacterium tuberculosis in NPA. The automated workstation yields nucleic acid with comparable extraction efficiency to manual protocols, which include commercially-available Qiagen spin column kits, across each of these sample types. This work expands the scope of applications beyond previous reports of TruTip to include difficult-to-disrupt cell types and automates the process, including a method for removal of organics, inside a compact bench-top workstation.
•A new lysis and homogenization method is combined with nucleic acid purification.•Nucleic acid purification and concentration occur in a pipette tip.•Both blood and nasopharyngeal samples are processed on a new Workstation.•Nucleic acid extraction from bacteria and viruses are automated on the Workstation.•The automated Workstation shows comparable recoveries to standard manual methods. |
doi_str_mv | 10.1016/j.mimet.2018.03.021 |
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•A new lysis and homogenization method is combined with nucleic acid purification.•Nucleic acid purification and concentration occur in a pipette tip.•Both blood and nasopharyngeal samples are processed on a new Workstation.•Nucleic acid extraction from bacteria and viruses are automated on the Workstation.•The automated Workstation shows comparable recoveries to standard manual methods.</description><identifier>ISSN: 0167-7012</identifier><identifier>EISSN: 1872-8359</identifier><identifier>DOI: 10.1016/j.mimet.2018.03.021</identifier><identifier>PMID: 29678500</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Automation, Laboratory - instrumentation ; Automation, Laboratory - methods ; Blood Cells - chemistry ; Complex Mixtures - isolation & purification ; Extraction ; Humans ; Influenza A virus - chemistry ; Methicillin-Resistant Staphylococcus aureus - chemistry ; Molecular Diagnostic Techniques - instrumentation ; Molecular Diagnostic Techniques - methods ; Mycobacterium tuberculosis - chemistry ; Nucleic acid isolation ; Nucleic Acids - isolation & purification ; Sample preparation ; Specimen Handling - instrumentation ; Specimen Handling - methods</subject><ispartof>Journal of microbiological methods, 2018-05, Vol.148, p.174-180</ispartof><rights>2018 Elsevier B.V.</rights><rights>Copyright © 2018 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c459t-9976a4b1b83e31947f6688745024fe21556c3772baa49f06783a6ea22f56be0a3</citedby><cites>FETCH-LOGICAL-c459t-9976a4b1b83e31947f6688745024fe21556c3772baa49f06783a6ea22f56be0a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0167701218300058$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,776,780,881,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29678500$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Thakore, Nitu</creatorcontrib><creatorcontrib>Garber, Steve</creatorcontrib><creatorcontrib>Bueno, Arial</creatorcontrib><creatorcontrib>Qu, Peter</creatorcontrib><creatorcontrib>Norville, Ryan</creatorcontrib><creatorcontrib>Villanueva, Michael</creatorcontrib><creatorcontrib>Chandler, Darrell P.</creatorcontrib><creatorcontrib>Holmberg, Rebecca</creatorcontrib><creatorcontrib>Cooney, Christopher G.</creatorcontrib><title>A bench-top automated workstation for nucleic acid isolation from clinical sample types</title><title>Journal of microbiological methods</title><addtitle>J Microbiol Methods</addtitle><description>Systems that automate extraction of nucleic acid from cells or viruses in complex clinical matrices have tremendous value even in the absence of an integrated downstream detector. We describe our bench-top automated workstation that integrates our previously-reported extraction method – TruTip – with our newly-developed mechanical lysis method. This is the first report of this method for homogenizing viscous and heterogeneous samples and lysing difficult-to-disrupt cells using "MagVor": a rotating magnet that rotates a miniature stir disk amidst glass beads confined inside of a disposable tube. Using this system, we demonstrate automated nucleic acid extraction from methicillin-resistant Staphylococcus aureus (MRSA) in nasopharyngeal aspirate (NPA), influenza A in nasopharyngeal swabs (NPS), human genomic DNA from whole blood, and Mycobacterium tuberculosis in NPA. The automated workstation yields nucleic acid with comparable extraction efficiency to manual protocols, which include commercially-available Qiagen spin column kits, across each of these sample types. This work expands the scope of applications beyond previous reports of TruTip to include difficult-to-disrupt cell types and automates the process, including a method for removal of organics, inside a compact bench-top workstation.
•A new lysis and homogenization method is combined with nucleic acid purification.•Nucleic acid purification and concentration occur in a pipette tip.•Both blood and nasopharyngeal samples are processed on a new Workstation.•Nucleic acid extraction from bacteria and viruses are automated on the Workstation.•The automated Workstation shows comparable recoveries to standard manual methods.</description><subject>Automation, Laboratory - instrumentation</subject><subject>Automation, Laboratory - methods</subject><subject>Blood Cells - chemistry</subject><subject>Complex Mixtures - isolation & purification</subject><subject>Extraction</subject><subject>Humans</subject><subject>Influenza A virus - chemistry</subject><subject>Methicillin-Resistant Staphylococcus aureus - chemistry</subject><subject>Molecular Diagnostic Techniques - instrumentation</subject><subject>Molecular Diagnostic Techniques - methods</subject><subject>Mycobacterium tuberculosis - chemistry</subject><subject>Nucleic acid isolation</subject><subject>Nucleic Acids - isolation & purification</subject><subject>Sample preparation</subject><subject>Specimen Handling - instrumentation</subject><subject>Specimen Handling - methods</subject><issn>0167-7012</issn><issn>1872-8359</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc1u1TAQRi0EopfCEyAhL9kk-Dd2FiBVFQWkSmxALC3HmVBfnDjYTlHfvm7vpYJNV7OYM9-M5iD0mpKWEtq927ezn6G0jFDdEt4SRp-gHdWKNZrL_inaVUo1ilB2gl7kvCeESi70c3TC-k5pScgO_TjDAyzuqilxxXYrcbYFRvwnpl-52OLjgqeY8LK5AN5h6_yIfY7h2Epxxi74xTsbcLbzGgCXmxXyS_RssiHDq2M9Rd8vPn47_9xcfv305fzssnFC9qXpe9VZMdBBc-C0F2rqOq2VkISJCRiVsnNcKTZYK_qJ1Ku57cAyNsluAGL5KfpwyF23YYbRwVKSDWZNfrbpxkTrzf-dxV-Zn_HayF4ILVUNeHsMSPH3BrmY2WcHIdgF4pYNI0z3kinJKsoPqEsx5wTTwxpKzJ0Sszf3SsydEkO4qUrq1Jt_L3yY-eugAu8PANQ_XXtIJjtfncDoE7hixugfXXALEcSfew</recordid><startdate>20180501</startdate><enddate>20180501</enddate><creator>Thakore, Nitu</creator><creator>Garber, Steve</creator><creator>Bueno, Arial</creator><creator>Qu, Peter</creator><creator>Norville, Ryan</creator><creator>Villanueva, Michael</creator><creator>Chandler, Darrell P.</creator><creator>Holmberg, Rebecca</creator><creator>Cooney, Christopher G.</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20180501</creationdate><title>A bench-top automated workstation for nucleic acid isolation from clinical sample types</title><author>Thakore, Nitu ; Garber, Steve ; Bueno, Arial ; Qu, Peter ; Norville, Ryan ; Villanueva, Michael ; Chandler, Darrell P. ; Holmberg, Rebecca ; Cooney, Christopher G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c459t-9976a4b1b83e31947f6688745024fe21556c3772baa49f06783a6ea22f56be0a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Automation, Laboratory - instrumentation</topic><topic>Automation, Laboratory - methods</topic><topic>Blood Cells - chemistry</topic><topic>Complex Mixtures - isolation & purification</topic><topic>Extraction</topic><topic>Humans</topic><topic>Influenza A virus - chemistry</topic><topic>Methicillin-Resistant Staphylococcus aureus - chemistry</topic><topic>Molecular Diagnostic Techniques - instrumentation</topic><topic>Molecular Diagnostic Techniques - methods</topic><topic>Mycobacterium tuberculosis - chemistry</topic><topic>Nucleic acid isolation</topic><topic>Nucleic Acids - isolation & purification</topic><topic>Sample preparation</topic><topic>Specimen Handling - instrumentation</topic><topic>Specimen Handling - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Thakore, Nitu</creatorcontrib><creatorcontrib>Garber, Steve</creatorcontrib><creatorcontrib>Bueno, Arial</creatorcontrib><creatorcontrib>Qu, Peter</creatorcontrib><creatorcontrib>Norville, Ryan</creatorcontrib><creatorcontrib>Villanueva, Michael</creatorcontrib><creatorcontrib>Chandler, Darrell P.</creatorcontrib><creatorcontrib>Holmberg, Rebecca</creatorcontrib><creatorcontrib>Cooney, Christopher G.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of microbiological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Thakore, Nitu</au><au>Garber, Steve</au><au>Bueno, Arial</au><au>Qu, Peter</au><au>Norville, Ryan</au><au>Villanueva, Michael</au><au>Chandler, Darrell P.</au><au>Holmberg, Rebecca</au><au>Cooney, Christopher G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A bench-top automated workstation for nucleic acid isolation from clinical sample types</atitle><jtitle>Journal of microbiological methods</jtitle><addtitle>J Microbiol Methods</addtitle><date>2018-05-01</date><risdate>2018</risdate><volume>148</volume><spage>174</spage><epage>180</epage><pages>174-180</pages><issn>0167-7012</issn><eissn>1872-8359</eissn><abstract>Systems that automate extraction of nucleic acid from cells or viruses in complex clinical matrices have tremendous value even in the absence of an integrated downstream detector. We describe our bench-top automated workstation that integrates our previously-reported extraction method – TruTip – with our newly-developed mechanical lysis method. This is the first report of this method for homogenizing viscous and heterogeneous samples and lysing difficult-to-disrupt cells using "MagVor": a rotating magnet that rotates a miniature stir disk amidst glass beads confined inside of a disposable tube. Using this system, we demonstrate automated nucleic acid extraction from methicillin-resistant Staphylococcus aureus (MRSA) in nasopharyngeal aspirate (NPA), influenza A in nasopharyngeal swabs (NPS), human genomic DNA from whole blood, and Mycobacterium tuberculosis in NPA. The automated workstation yields nucleic acid with comparable extraction efficiency to manual protocols, which include commercially-available Qiagen spin column kits, across each of these sample types. This work expands the scope of applications beyond previous reports of TruTip to include difficult-to-disrupt cell types and automates the process, including a method for removal of organics, inside a compact bench-top workstation.
•A new lysis and homogenization method is combined with nucleic acid purification.•Nucleic acid purification and concentration occur in a pipette tip.•Both blood and nasopharyngeal samples are processed on a new Workstation.•Nucleic acid extraction from bacteria and viruses are automated on the Workstation.•The automated Workstation shows comparable recoveries to standard manual methods.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>29678500</pmid><doi>10.1016/j.mimet.2018.03.021</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Automation, Laboratory - instrumentation Automation, Laboratory - methods Blood Cells - chemistry Complex Mixtures - isolation & purification Extraction Humans Influenza A virus - chemistry Methicillin-Resistant Staphylococcus aureus - chemistry Molecular Diagnostic Techniques - instrumentation Molecular Diagnostic Techniques - methods Mycobacterium tuberculosis - chemistry Nucleic acid isolation Nucleic Acids - isolation & purification Sample preparation Specimen Handling - instrumentation Specimen Handling - methods |
title | A bench-top automated workstation for nucleic acid isolation from clinical sample types |
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