title: A single-tube nucleic acid extraction, amplification, and detection method using aluminum oxide.
abstract: A disposable 0.2-ml polymerase chain reaction (PCR) tube modified with an aluminum oxide membrane (AOM) has been developed for the extraction, amplification, and detection of nucleic acids. To assess the dynamic range of AOM tubes for real-time PCR, quantified herpes simplex virus (HSV) DNA was used to compare AOM tubes to standard PCR tubes. AOM PCR tubes used for amplification and detection of quantified HSV-1 displayed a crossing threshold (C(T)) shift 0.1 cycles greater than PCR tube controls. Experiments with HSV-1-positive cerebrospinal fluid (CSF) examined the extraction, amplification, and detection properties of the AOM tubes compared to the Qiagen DNA blood mini kit. The AOM extraction, amplification, and detection of HSV-1 in CSF displayed differences of less than one C(T) when compared to Qiagen-extracted samples. Experiments testing the AOM method using clinical CSF samples displayed 100% concordance with reported results. AOM tubes have no adverse effects on amplification or fluorescence acquisition by real-time PCR and can be effectively used for the extraction, amplification, and detection of HSV from CSF. The AOM single tube method is a fast, reliable, and reproducible technique for the extraction, amplification, and detection of HSV in CSF.
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT 84108, USA. shale.dames@aruplab.com
authors.forename: Shale
authors.initials: S
authors.lastname: Dames
authors.forename: L Kathryn
authors.initials: LK
authors.lastname: Bromley
authors.forename: Mark
authors.initials: M
authors.lastname: Herrmann
authors.forename: Marc
authors.initials: M
authors.lastname: Elgort
authors.forename: Maria
authors.initials: M
authors.lastname: Erali
authors.forename: Roger
authors.initials: R
authors.lastname: Smith
authors.forename: Karl V
authors.initials: KV
authors.lastname: Voelkerding
mesh_terms: D000537:Aluminum Oxide / Q000737:chemistry*; D004279:DNA, Viral / Q000134:cerebrospinal fluid*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation*; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D011126:Polypropylenes; D018139:Simplexvirus / Q000378:metabolism
chemical_list: D004279:DNA, Viral; D011126:Polypropylenes; D000537:Aluminum Oxide