Quantification of HIV-1 DNA using Real-Time Recombinase Polymerase Amplification

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dc.citation.firstpage5615
dc.citation.issueNumber12
dc.citation.journalTitleAnalytical Chemistry
dc.citation.lastpage5619
dc.citation.volumeNumber86
dc.contributor.authorCrannell, Zachary Austin
dc.contributor.authorRohrman, Brittany
dc.contributor.authorRichards-Kortum, Rebecca
dc.date.accessioned2014-10-29T18:15:02Z
dc.date.available2014-10-29T18:15:02Z
dc.date.issued2014
dc.description.abstractAlthough recombinase polymerase amplification (RPA) has many advantages for the detection of pathogenic nucleic acids in point-of-care applications, RPA has not yet been applied for has not yet been implemented to quantify sample concentration using a standard curve. Here we describe a real-time RPA assay with an internal positive control and an algorithm that analyzes the real-time fluorescence data to quantify HIV-1 DNA. We show that DNA concentration and the onset of detectable amplification are correlated by an exponential standard curve. In a set of experiments in which the standard curve and algorithm were used to analyze and quantify additional DNA samples, the algorithm predicted an average concentration within one order of magnitude of the correct concentration for all HIV-1 DNA concentrations tested. These results suggest that qRPA may serve as a powerful tool for quantifying nucleic acids and may be adapted for use in single-sample point-of-care diagnostic systems.
dc.identifier.citationCrannell, Zachary Austin, Rohrman, Brittany and Richards-Kortum, Rebecca. "Quantification of HIV-1 DNA using Real-Time Recombinase Polymerase Amplification." <i>Analytical Chemistry,</i> 86, no. 12 (2014) American Chemical Society: 5615-5619. http://dx.doi.org/10.1021/ac5011298.
dc.identifier.doihttp://dx.doi.org/10.1021/ac5011298
dc.identifier.urihttps://hdl.handle.net/1911/77665
dc.language.isoeng
dc.publisherAmerican Chemical Society
dc.rightsThis is an author's peer-reviewed final manuscript, as accepted by the publisher. The published article is copyrighted by the American Chemical Society.
dc.titleQuantification of HIV-1 DNA using Real-Time Recombinase Polymerase Amplification
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpost-print
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