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A bench-top automated workstation for nucleic acid isolation from clinical sample types.
Thakore, Nitu; Garber, Steve; Bueno, Arial; Qu, Peter; Norville, Ryan; Villanueva, Michael; Chandler, Darrell P; Holmberg, Rebecca; Cooney, Christopher G.
Afiliación
  • Thakore N; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States.
  • Garber S; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States.
  • Bueno A; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States.
  • Qu P; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States.
  • Norville R; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States.
  • Villanueva M; Laboratorios Medicos Especializados, Ave Ejercito Nacional #6008-9, Partido Escobedo, Cd. Juarez, CHIH 32320, Mexico.
  • Chandler DP; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States.
  • Holmberg R; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States.
  • Cooney CG; Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick, MD 21701, United States. Electronic address: cooney@akonni.com.
J Microbiol Methods ; 148: 174-180, 2018 05.
Article en En | MEDLINE | ID: mdl-29678500
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Manejo de Especímenes / Ácidos Nucleicos / Técnicas de Diagnóstico Molecular / Automatización de Laboratorios Tipo de estudio: Diagnostic_studies / Guideline Límite: Humans Idioma: En Revista: J Microbiol Methods Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Países Bajos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Manejo de Especímenes / Ácidos Nucleicos / Técnicas de Diagnóstico Molecular / Automatización de Laboratorios Tipo de estudio: Diagnostic_studies / Guideline Límite: Humans Idioma: En Revista: J Microbiol Methods Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Países Bajos