Bronze M., Aitken S.C., Wallis C.L., Steegen K., Stuyver L.J., de Wit T.F.R., Stevens W.
Department of Molecular Medicine and Haematology, University of the Witwatersrand, Johannesburg, South Africa; Department of Molecular Medicine and Haematology, National Health Laboratory Services, Charlotte Maxeke Johannesburg Academic Hospital, South Africa; University Medical Centre Utrecht (UMCU), Netherlands; Janssen Diagnostics, Beerse, Belgium; Department of Global Health, Academic Medical Center, University of Amsterdam, Amsterdam Institute for Global Health and Development, Netherlands
Bronze, M., Department of Molecular Medicine and Haematology, University of the Witwatersrand, Johannesburg, South Africa, Department of Molecular Medicine and Haematology, National Health Laboratory Services, Charlotte Maxeke Johannesburg Academic Hospital, South Africa; Aitken, S.C., University Medical Centre Utrecht (UMCU), Netherlands; Wallis, C.L., Department of Molecular Medicine and Haematology, University of the Witwatersrand, Johannesburg, South Africa; Steegen, K., Department of Molecular Medicine and Haematology, University of the Witwatersrand, Johannesburg, South Africa; Stuyver, L.J., Janssen Diagnostics, Beerse, Belgium; de Wit, T.F.R., Department of Global Health, Academic Medical Center, University of Amsterdam, Amsterdam Institute for Global Health and Development, Netherlands; Stevens, W., Department of Molecular Medicine and Haematology, University of the Witwatersrand, Johannesburg, South Africa, Department of Molecular Medicine and Haematology, National Health Laboratory Services, Charlotte Maxeke Johannesburg Academic Hospital, South Africa
HIV-1 RNA viral load is the preferred tool to monitor virological failure during antiretroviral therapy (ART) exposure. Timely detection of virological failure can reduce the prevalence and complexity of HIV-1 drug resistance. This field evaluation further characterizes a two-step approach to identify virological failure, as a measure of ART adherence, and detect HIVDR mutations in the reverse transcriptase (RT) gene of HIV-1. Two hundred and forty-eight (248) samples were tested; 225 from South African HIV-1 participants enrolled in the PharmAccess African Studies to Evaluate Resistance (PASER) cohort, forty of which had paired dried blood spot (DBS) samples and 23 HIV-1 negative samples. A newly developed virological failure assay (ARTA-VFA) was used on all samples, and those with a viral load >5000 RNA copies/ml were genotyped with a shortened RT protocol to detect HIVDR (ARTA-HIVDRultralight). The ARTA-VFA showed good precision and linearity as compared to a commercial reference assay (NucliSENS® EasyQ v1.2, Roche) with an R2 of 0.99. Accuracy studies illustrated standard deviations of <1 log RNA copies/ml for plasma and DBS ARTA-VFA results compared to the reference method. The ARTA-VFA's intended use was to deliver qualitative results either < or >5000 RNA copies/ml. No significant differences in the proportion of results < or > either the 5000 RNA copies/ml or 1000 RNA copies/ml cut-off were noted for plasma indicating either cut-off to be useful. Significant differences were noted in these proportions when DBS were used (P=0.0002), where a 5000 RNA copies/ml cut-off was deemed more appropriate. The sensitivity and specificity of the ARTA-VFA with plasma were 95% and 93% and 91% and 95% for DBS using a 5000 RNA copies/ml cut-off. The ARTA HIVDRultralight assay was reliable for plasma and DBS samples with a viral load >5000 RNA copies/ml, with amplification and sequencing success rates of 91% and 92% respectively for plasma, and 95% and 80% respectively for DBS. HIVDR profiles for plasma and DBS were 100% concordant with the reference assay. This study evaluated a previously described combination of two assays potentially useful in assessing HIV-1 virological failure and resistance, showing good concordance with reference assays. These assays are simple to perform and are affordable, viable options to detect virological failures in certain resource limited settings. The assays' compatibility with DBS sampling extends the access of HIV-1 virological monitoring to more remote settings. © 2013 Elsevier B.V.
antiretrovirus agent; accuracy; antiviral resistance; article; controlled study; cost; diagnostic kit; dried blood spot testing; drug treatment failure; genotyping technique; human; Human immunodeficiency virus 1; intermethod comparison; nonhuman; predictive value; priority journal; real time polymerase chain reaction; sensitivity and specificity; sequence analysis; virological failure assay; virus examination; virus load; Antiretroviral drug resistance; Genotyping; HIV-1 virological failure assay; Anti-Retroviral Agents; Drug Resistance, Viral; HIV Infections; HIV-1; Humans; Microbial Sensitivity Tests; Molecular Diagnostic Techniques; Sensitivity and Specificity; South Africa; Treatment Failure; Viral Load
