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
Hauptverfasser: Thakore, Nitu, Garber, Steve, Bueno, Arial, Qu, Peter, Norville, Ryan, Villanueva, Michael, Chandler, Darrell P., Holmberg, Rebecca, Cooney, Christopher G.
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container_end_page 180
container_issue
container_start_page 174
container_title Journal of microbiological methods
container_volume 148
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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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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