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Analytical performance of an automated assay quantifying HIV-1 from dried blood spots

, : Analytical performance of an automated assay quantifying HIV-1 from dried blood spots. Journal of Clinical Virology 57(3): 271-273

Performance of an automated sample preparation and viral load quantification for HIV-positive dried blood spots (DBS) on the Siemens VERSANT kPCR Molecular System has been previously demonstrated with clinical samples. Evaluation of the analytical performance of the automated assay using HIV-positive DBS prepared from a dilution series. Over 300 DBS of HIV-1 nucleic acids from a dilution series in lysed whole blood (322copies/mL to over 1.6 107 copies/mL) were spotted onto Whatman 903 cards and analyzed to evaluate analytical performance. Cross contamination was examined with a checkerboard pattern of 82 alternating negative and 1 106 copies/mL samples. Analytical sensitivity evaluation with a single 50?L spot demonstrated a limit of detection (LoD) of 866copies/mL. Above the LoD, linearity (difference between linearized and observed mean values) was within 0.10log, and accuracy (difference between expected and observed mean values) was within 0.18log. Imprecision for dilution series levels more than two-fold above the LoD was measured as 20% to 27% CV of quantification. No cross contamination was observed. The HIV-1 DBS Assay performed similarly to the VERSANT HIV-1 RNA 1.0 Assay (kPCR) in assay linearity, accuracy, and imprecision. The assay was sensitive enough to run single 50?L spots and used an unmodified VERSANT SP Module with only a 30min incubation prior to automated sample preparation. DBS-specific assay calibrators and controls, expressly formulated for easy frozen storage and identical processing to samples, were employed.


PMID: 23571052

DOI: 10.1016/j.jcv.2013.03.001

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