title: Quantification of HIV-1 DNA using real-time recombinase polymerase amplification.
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 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 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 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 acids and may be adapted for use in single-sample point-of-care diagnostic systems.
authors.affiliation: Rice University , 6500 Main Street, Houston, Texas 77030, United States.
authors.forename: Zachary Austin
authors.initials: ZA
authors.lastname: Crannell
authors.forename: Brittany
authors.initials: B
authors.lastname: Rohrman
authors.forename: Rebecca
authors.initials: R
authors.lastname: Richards-Kortum
mesh_terms: D000465:Algorithms; D004279:DNA, Viral / Q000032:analysis*; D015658:HIV Infections / Q000175:diagnosis; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D019095:Point-of-Care Systems; D045522:Recombinases / Q000378:metabolism*
chemical_list: D004279:DNA, Viral; D045522:Recombinases