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dc.contributor.authorNatoli, Mary E.
Rohrman, Brittany A.
De Santiago, Carolina
van Zyl, Gert U.
Richards-Kortum, Rebecca R.
dc.date.accessioned 2018-07-11T15:59:27Z
dc.date.available 2018-07-11T15:59:27Z
dc.date.issued 2018
dc.identifier.citation Natoli, Mary E., Rohrman, Brittany A., De Santiago, Carolina, et al.. "Paper-based detection of HIV-1 drug resistance using isothermal amplification and an oligonucleotide ligation assay." Analytical Biochemistry, 544, (2018) Elsevier: 64-71. https://doi.org/10.1016/j.ab.2017.12.008.
dc.identifier.urihttps://hdl.handle.net/1911/102373
dc.description.abstract Regular HIV-1 viral load monitoring is the standard of care to assess antiretroviral therapy effectiveness in resource-rich settings. Persistently elevated viral loads indicate virologic failure (VF), which warrants HIV drug resistance testing (HIVDRT) to allow individualized regimen switches. However, in settings lacking access to HIVDRT, clinical decisions are often made based on symptoms, leading to unnecessary therapy switches and increased costs of care. This work presents a proof-of-concept assay to detect M184V, the most common drug resistance mutation after first-line antiretroviral therapy failure, in a paper format. The first step isothermally amplifies a section of HIV-1ᅠreverse transcriptaseᅠcontaining M184V using a recombinase polymerase amplification (RPA) assay. Then, an oligonucleotide ligation assay (OLA) is used to selectively label the mutant and wild type amplified sequences. Finally, a lateral flow enzyme-linked immunosorbent assay (ELISA) differentiates between OLA-labeled products with or without M184V. Our method shows 100% specificity and 100% sensitivity when tested with samples that contained 200 copies of mutant DNA and 800 copies of wild type DNA prior to amplification. When integrated with sample preparation, this method may detect HIV-1 drug resistance at a low cost and at a rural hospital laboratory.
dc.language.iso eng
dc.publisher Elsevier
dc.rightsThis is an open access article under the CC BY license (http://creativecommons.org/licenses/BY/4.0/).
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.title Paper-based detection of HIV-1 drug resistance using isothermal amplification and an oligonucleotide ligation assay
dc.type Journal article
dc.citation.journalTitle Analytical Biochemistry
dc.subject.keywordrecombinase polymerase amplification
HIV drug resistance
lateral flow
oligonucleotide ligation assay
dc.citation.volumeNumber 544
dc.type.dcmi Text
dc.identifier.doihttps://doi.org/10.1016/j.ab.2017.12.008
dc.identifier.pmcid PMC5854266
dc.identifier.pmid 29229373
dc.type.publication publisher version
dc.citation.firstpage 64
dc.citation.lastpage 71


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This is an open access article under the CC BY license (http://creativecommons.org/licenses/BY/4.0/).
Except where otherwise noted, this item's license is described as This is an open access article under the CC BY license (http://creativecommons.org/licenses/BY/4.0/).