Paper-based detection of HIV-1 drug resistance using isothermal amplification and an oligonucleotide ligation assay

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dc.citation.firstpage64en_US
dc.citation.journalTitleAnalytical Biochemistryen_US
dc.citation.lastpage71en_US
dc.citation.volumeNumber544en_US
dc.contributor.authorNatoli, Mary E.en_US
dc.contributor.authorRohrman, Brittany A.en_US
dc.contributor.authorDe Santiago, Carolinaen_US
dc.contributor.authorvan Zyl, Gert U.en_US
dc.contributor.authorRichards-Kortum, Rebecca R.en_US
dc.contributor.orgBioengineeringen_US
dc.date.accessioned2018-07-11T15:59:27Zen_US
dc.date.available2018-07-11T15:59:27Zen_US
dc.date.issued2018en_US
dc.description.abstractRegular 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-1ļ¾ reverse transcriptaseļ¾ containing 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.en_US
dc.identifier.citationNatoli, 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." <i>Analytical Biochemistry,</i> 544, (2018) Elsevier: 64-71. https://doi.org/10.1016/j.ab.2017.12.008.en_US
dc.identifier.doihttps://doi.org/10.1016/j.ab.2017.12.008en_US
dc.identifier.urihttps://hdl.handle.net/1911/102373en_US
dc.language.isoengen_US
dc.publisherElsevieren_US
dc.rightsThis is an open access article under the CC BY license (http://creativecommons.org/licenses/BY/4.0/).en_US
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en_US
dc.subject.keywordrecombinase polymerase amplificationen_US
dc.subject.keywordHIV drug resistanceen_US
dc.subject.keywordlateral flowen_US
dc.subject.keywordoligonucleotide ligation assayen_US
dc.titlePaper-based detection of HIV-1 drug resistance using isothermal amplification and an oligonucleotide ligation assayen_US
dc.typeJournal articleen_US
dc.type.dcmiTexten_US
dc.type.publicationpublisher versionen_US
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