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dc.contributor.authorRohrman, Brittany
Richards-Kortum, Rebecca
dc.date.accessioned 2015-03-19T15:07:13Z
dc.date.available 2015-03-19T15:07:13Z
dc.date.issued 2015
dc.identifier.citation Rohrman, Brittany and Richards-Kortum, Rebecca. "Inhibition of Recombinase Polymerase Amplification by Background DNA: A Lateral Flow-Based Method for Enriching Target DNA." Analytical Chemistry, 87, no. 3 (2015) American Chemical Society: 1963-1967. http://dx.doi.org/10.1021/ac504365v.
dc.identifier.urihttps://hdl.handle.net/1911/79391
dc.description.abstract Recombinase polymerase amplification (RPA) may be used to detect a variety of pathogens, often after minimal sample preparation. However, previous work has shown that whole blood inhibits RPA. In this paper, we show that the concentrations of background DNA found in whole blood prevent the amplification of target DNA by RPA. First, using an HIV-1 RPA assay with known concentrations of nonspecific background DNA, we show that RPA tolerates more background DNA when higher HIV-1 target concentrations are present. Then, using three additional assays, we demonstrate that the maximum amount of background DNA that may be tolerated in RPA reactions depends on the DNA sequences used in the assay. We also show that changing the RPA reaction conditions, such as incubation time and primer concentration, has little effect on the ability of RPA to function when high concentrations of background DNA are present. Finally, we develop and characterize a lateral flow-based method for enriching the target DNA concentration relative to the background DNA concentration. This sample processing method enables RPA of 104 copies of HIV-1 DNA in a background of 0–14 μg of background DNA. Without lateral flow sample enrichment, the maximum amount of background DNA tolerated is 2 μg when 106 copies of HIV-1 DNA are present. This method requires no heating or other external equipment, may be integrated with upstream DNA extraction and purification processes, is compatible with the components of lysed blood, and has the potential to detect HIV-1 DNA in infant whole blood with high proviral loads.
dc.language.iso eng
dc.publisher American Chemical Society
dc.rights Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
dc.title Inhibition of Recombinase Polymerase Amplification by Background DNA: A Lateral Flow-Based Method for Enriching Target DNA
dc.type Journal article
dc.citation.journalTitle Analytical Chemistry
dc.citation.volumeNumber 87
dc.citation.issueNumber 3
dc.type.dcmi Text
dc.identifier.doihttp://dx.doi.org/10.1021/ac504365v
dc.identifier.pmid 25560368
dc.type.publication publisher version
dc.citation.firstpage 1963
dc.citation.lastpage 1967


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