Both GTT and PTT rely on the initial amplification of the HIV-1 glycoprotein 160 (gp160) or gp120 coding sequence from plasma viral RNA. A minimal amount of HIV-1 RNA (>500 copies/mL) is needed for successful amplification. In many patients with early failure for whom a treatment change is considered, the HIV-1 RNA will not reach this level. In addition, for some patients a treatment change may be considered when the viral load is suppressed, for example to address problems of toxicity. Proviral selleck screening library DNA may be considered a potential alternative source of viral genetic material for tropism testing in patients with low or undetectable

viral load. Cellular proviral DNA contains the reservoir of archived viruses, and it has been shown that V3 sequences predicted to derive from X4 viruses are present and even enriched in this reservoir [10–12]. The aim of this study was to evaluate GTT on plasma RNA and proviral DNA for two groups

of patients. The first group comprised treated and untreated patients with a viral load of >500 copies/mL who underwent parallel testing of plasma RNA and proviral DNA. For the majority of these samples, results of PTT were also available, obtained through the use of either the MT2 assay or the Trofile™ assays (Monogram). The second group comprised treated patients with a viral load of <500 copies/mL who underwent GTT on a current proviral DNA sample and on the last stored plasma RNA sample collected before the viral load dropped to undetectable levels because of ART initiation. Blood samples were collected at five AIDS Reference Centres in Belgium and Luxembourg, and at the Royal selleck Free Hospital in London, UK. A first series, named ‘simultaneous RNA/DNA’, consisted of plasma and blood cell samples collected on the same day from 220 patients with a viral load of >500 copies/mL. Of these 220 patients, 101 were treatment-naïve and 119 were treatment-experienced. Results of PTT were available for 142 individuals, after performing the MT2 assay (n=72), the original Trofile™ assay (OTA; Monogram) (n=24) or the enhanced Trofile™

assay (ESTA; Monogram) (n=46). A second series of samples, named ‘longitudinal RNA/DNA’, was collected from 137 individuals with a viral load of <500 copies/mL. GTT was performed on a current proviral DNA sample and on a stored Florfenicol plasma RNA sample with a viral load of >500 copies/mL, collected shortly before starting or adapting ART. At the time of plasma sample collection, 108 patients were treatment-naïve, 20 had temporarily interrupted their ART and nine were on a failing regimen. The subtype distribution of selected samples was 67.6% B vs. 32.4% non-B. Samples were collected with informed consent and the study was conducted with the approval of the ethics committees of the participating institutions. Plasma and buffy coat cells were separated from ethylenediaminetetraacetic acid (EDTA)-anti-coagulated blood and stored frozen at −80 °C until analysis.