JMD Association for Molecular Pathology 2008 Annual Meeting
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JMD 2006, Vol. 8, No. 1
Copyright © 2006 American Society for Investigative Pathology & Association for Molecular Pathology

Technical Advances

A Single-Tube Nucleic Acid Extraction, Amplification, and Detection Method Using Aluminum Oxide

Shale Dames*, L. Kathryn Bromley*, Mark Herrmann*, Marc Elgort*, Maria Erali*, Roger Smith*,{dagger} and Karl V. Voelkerding*,{dagger}

From the ARUP Institute for Clinical and Experimental Pathology, * Salt Lake City; and the Department of Pathology, {dagger} University of Utah, Salt Lake City, Utah

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 (CT) 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 CT 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.






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Copyright © 2006 by the American Society for Investigative Pathology and the Association for Molecular Pathology.