Booth, C.

Garcia-Diaz, A.

Johnson, M.

Macartney, M.

McCormick, A.L.

Webster, D.

Youle, M.

English

WestminsterResearch

ORAL PRESENTATION Open AccessFeasibility of testing and detection of HIV-1 drugresistance in proviral DNAClare L Booth*, Adele McCormick, Ana Garcia-Diaz, Malcolm Macartney, Michael Youle, Margaret Johnson,Daniel WebsterFrom The 7th Romanian National HIV/AIDS Congress and The 2nd Central European HIV ForumSibiu, Romania. 29-31 May 2014In recent years antiretroviral availability and access hasbecome more prevalent in resource poor settings, whilstthe availability of access to HIV-1 drug resistance testingremains limited and patients are often started on therapyor switched when failing without a resistance test. ProviralDNA has been proposed as a source of archived resistancemutations. We investigated the feasibility of using proviralDNA to detect previously documented plasma resistancemutations.Amplification and sequencing of pol from proviral DNAderived from buffy coat (BCL), buffy coat dried on filterpaper (BCS) or dried blood spots (DBS) was performedusing an in-house assay.Only 2/20 samples were amplified from long term storedbuffy coat (>1 year) compared to at least one prospectivesample from 10/20 patients. Six patients had a plasmaviral load (PVL) <40 cp/ml at time of sampling, though4/6 only become undetectable on the most recent sample,2/4 had been undetectable for 149 and 819 days respec-tively. The remaining 6 patients had detectable PVL(median 3.1 log10 cp/ml, range: 1.7–6.2 log10 cp/ml). Incontrast, 21/28 (75%) of samples which failed to amplifywere <40 cp/ml, 2 had become undetectable in the mostrecent sample and 19 had been undetectable for a medianof 577 days (range: 32–2481 days).6/10 BCL samples showed none of the previouslydocumented resistance for those patients, 3/10 showedsome of the previously known mutations and only 1/10showed all historic resistance, though this patient hadonly one documented mutation (K103N). For the BCSsamples 3/4 showed none of the documented resistance,1/4 harboured the previously known mutations (M184V,K103N). 3/6 of the DBS had none of the previouslydocumented resistance, 2/6 showed some of the knownmutations and 1/6 showed both known resistance muta-tions (M41L, T215S). In patients where multiple specimentypes were amplified, 3/5 showed differences in mutationsdetected between sample types, with 2/5 correlating buthaving no mutations detected in any specimens.In patients with undetectable PVL, particularly thoseundetectable for long periods, it was difficult to sequencearchived DNA. Concordance between different specimentypes was variable as was concordance with previouslydocumented mutations. Historic resistance reportsremain important in the clinical management of patientson antiretroviral therapy, though proviral DNA testingmay be useful in patients where historic reports are notavailable.Published: 29 May 2014doi:10.1186/1471-2334-14-S4-O25Cite this article as: Booth et al.: Feasibility of testing and detection ofHIV-1 drug resistance in proviral DNA. BMC Infectious Diseases 201414(Suppl 4):O25.Submit your next manuscript to BioMed Centraland take full advantage of: • Convenient online submission• Thorough peer review• No space constraints or color figure charges• Immediate publication on acceptance• Inclusion in PubMed, CAS, Scopus and Google Scholar• Research which is freely available for redistributionSubmit your manuscript at www.biomedcentral.com/submitRoyal Free London NHS Foundation Trust, London, United KingdomBooth et al. BMC Infectious Diseases 2014, 14(Suppl 4):O25http://www.biomedcentral.com/1471-2334/14/S4/O25© 2014 Booth et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative CommonsAttribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction inany medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Feasibility of testing and detection of HIV-1 drug resistance in proviral DNA

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Feasibility of testing and detection of HIV-1 drug resistance in proviral DNA

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