| Literature DB >> 29678500 |
Nitu Thakore1, Steve Garber1, Arial Bueno1, Peter Qu1, Ryan Norville1, Michael Villanueva2, Darrell P Chandler1, Rebecca Holmberg1, Christopher G Cooney3.
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.Entities:
Keywords: Extraction; Nucleic acid isolation; Sample preparation
Mesh:
Substances:
Year: 2018 PMID: 29678500 PMCID: PMC5944857 DOI: 10.1016/j.mimet.2018.03.021
Source DB: PubMed Journal: J Microbiol Methods ISSN: 0167-7012 Impact factor: 2.363