Validation for clinical use of a novel HIV-2 plasma RNA viral load assay using the Abbott m2000 platform

BackgroundOptimal care of persons infected with human immunodeficiency virus type 2 (HIV-2) requires an accurate assessment of HIV-2 plasma viral load (VL), but no clinically approved quantitative HIV-2 RNA VL assay exists.ObjectivesTo validate a novel quantitative HIV-2 RNA assay for clinical and research use.Study designThe Abbott m2000sp/rt platform was adapted for quantification of HIV-2 RNA in plasma. Amplification targeted a region of the long terminal repeat conserved in Group A and B HIV-2. Electron microscopy-counted-HIV-2 standards, the WHO/NIBSC HIV-2 International Standard and clinical specimens (N = 162) were used to determine the precision, sensitivity, specificity, linear range, accuracy, and clinical performance of the assay.ResultsThe quantitative linear range of the HIV-2 RNA assay was 10–1,000,000 copies/mL (R2 > 0.99), with a limit of detection of 8 copies/mL (95% CI, 5–18 copies/mL). The assay did not cross-react with HIV-1, and quantification of HIV-2 RNA was not affected by the presence of >5 log10 HIV-1 RNA copies/mL. The total standard deviation (SD) and intra- and inter-run SD were 0.095, 0.093 and 0.162, respectively, at nominal inputs of 3.7, 1.7 and 1.0 log10 HIV-2 RNA copies/mL. The HIV-2 WHO/NIBSC International Standard (1000 IU) was shown to contain 152 RNA copies/mL (95% CI 141–163). Overall, HIV-2 RNA was quantified at ≥10 copies/mL from 86 (53%) clinical specimens (median, 2.24 log10 copies/mL; range 10–16,870), and nine specimens (6%) had HIV-2 RNA detected at <10 copies/mL.ConclusionsWe developed and validated a highly sensitive HIV-2 VL assay that is suitable for clinical and research use.