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

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    Author
    Crannell, Zachary Austin; Rohrman, Brittany; Richards-Kortum, Rebecca
    Date
    2014
    Abstract
    Although 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.
    Citation
    Crannell, Zachary Austin, Rohrman, Brittany and Richards-Kortum, Rebecca. "Quantification of HIV-1 DNA using Real-Time Recombinase Polymerase Amplification." Analytical Chemistry, 86, no. 12 (2014) American Chemical Society: 5615-5619. http://dx.doi.org/10.1021/ac5011298.
    Published Version
    http://dx.doi.org/10.1021/ac5011298
    Type
    Journal article
    Publisher
    American Chemical Society
    Citable link to this page
    https://hdl.handle.net/1911/77665
    Rights
    This is an author's peer-reviewed final manuscript, as accepted by the publisher. The published article is copyrighted by the American Chemical Society.
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    • Bioengineering Publications [632]
    • Faculty Publications [4990]

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    Home | FAQ | Contact Us | Privacy Notice | Accessibility Statement
    Managed by the Digital Scholarship Services at Fondren Library, Rice University
    Physical Address: 6100 Main Street, Houston, Texas 77005
    Mailing Address: MS-44, P.O.BOX 1892, Houston, Texas 77251-1892
    Site Map