1. Quantification of HIV-1 DNA Using Real-Time Recombinase Polymerase Amplification.
- Author
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Crannell, Zachary Austin, Rohrman, Brittany, and Richards-Kortum, Rebecca
- Subjects
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NUCLEIC acids , *HIV , *ALGORITHMS , *RECOMBINASES , *DNA , *GENETICS - Abstract
Although recombinase polymerase amplification (RPA) has many advantages for the detection of pathogenic nucleic adds in point-of-care applications, RPA 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 realtime 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 1 order of magnitude of the correct concentration for all HIV-1 DNA concentrations tested. These results suggest that quantitative RPA (qRPA) may serve as a powerful tool for quantifying nucleic adds and may be adapted for use in single-sample point-of-care diagnostic systems. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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