To explore this possibility, we studied patients with lengthy exp

To explore this possibility, we studied patients with lengthy exposure to the three main antiretroviral drug classes who had experienced multiple treatment failures. We compared the results of genotypic drug resistance assays on cellular DNA at the time of suppressed viraemia with those of resistance genotyping of plasma

HIV-1 RNA at the time of past treatment failures. The patients had been enrolled in the Agence Nationale de Recherche sur le SIDA (ANRS) 138-intEgrase inhibitor MK_0518 to Avoid Subcutaneous Injections of EnfuviRtide (EASIER) randomized trial, which was designed to assess the switch from enfuvirtide to raltegravir among highly treatment-experienced HIV-1-infected patients with good virological control [8]. The selleck chemical ANRS 138-EASIER study was a 48-week noninferiority randomized multicentre trial assessing the efficacy and safety of a switch from enfuvirtide to raltegravir in highly treatment-experienced patients receiving a suppressive enfuvirtide-containing regimen [8]. The main inclusion criteria were (i) HIV-1-infected patients with failure on, or intolerance to, triple drug classes [nucleoside reverse transcriptase inhibitor (NRTI), nonnucleoside reverse transcriptase inhibitor (NNRTI) and protease inhibitor (PI)]; (ii) a stable enfuvirtide-based regimen for 3 months or more; and (iii) plasma HIV-1 RNA levels < 400

copies/mL for at least the last 3 months. One hundred and sixty-nine patients were enrolled in this trial. The results of all available HIV-1 RNA resistance tests performed on plasma during previous episodes of virological failure were collected from French virology laboratories belonging to the ANRS network Enzalutamide purchase and performing annual resistance genotyping quality controls [9]. Information on drug resistance mutations was analysed centrally. We constructed the ‘cumulative’ RNA genotype for each patient Orotidine 5′-phosphate decarboxylase by adding up all mutations found in previous genotypic tests for each drug class. HIV-1 DNA resistance genotyping was performed in a central laboratory prior to randomization. Viral DNA was extracted from 200 μL of frozen stored whole blood using an automatic nucleic

acid extractor (MagnaPure; Roche, Meylan, France). Reverse transcriptase–polymerase chain reaction (RT-PCR) and then nested PCR were used to amplify the reverse transcriptase (RT) and protease (PR) genes according to ANRS consensus methods (www.hivfrenchresistance.org). Population sequencing was performed on purified amplicons with the Taq Dye Deoxy Terminator cycle sequencing kits (Applied Biosytems, Courtaboeuf, France) and resolved on an ABI 3700 automated DNA sequencer (Applied Biosytems, Courtaboeuf, France). Sequences were aligned with the HIV-1 subtype B HXB2 reference strain, and RT and PR mutations were identified from the 2008 International Antiviral Society (IAS)-USA resistance list (www.iasusa.org) and the 2009 ANRS v18 algorithm (www.hivfrenchresistance.org).

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