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.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Thakore N,Garber S,Bueno A,Qu P,Norville R,Villanueva M,Chandler DP,Holmberg R,Cooney CGdoi
10.1016/j.mimet.2018.03.021subject
Has Abstractpub_date
2018-05-01 00:00:00pages
174-180eissn
0167-7012issn
1872-8359pii
S0167-7012(18)30005-8journal_volume
148pub_type
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