[Analytical Performance of the ELITe BeGenius System for HIV-1 RNA and HCV RNA Quantification: Ensuring Diagnostic...

Quantitative detection of human immunodeficiency virus-type 1 (HIV-1) and hepatitis C virus (HCV) RNA plays a crucial role in the diagnosis, monitoring of the therapy and evaluation of the treatment response. The ELITe BeGenius® platform (ELITechGroup, Turin, Italy) is a fully automated sample-toresult molecular system integrating extraction, amplification and detection within a single workflow. The HIV-1 ELITe MGB® and HCV ELITe MGB® assays are real-time polymerase chain reaction tests designed for plasma viral-load quantification. This study aimed to verify their analytical performance under routine clinical laboratory conditions. Verification included assessments of accuracy, intra- and inter-assay precision, linearity and method correlation. A total of 70 plasma samples for HIV-1 RNA and 52 for HCV RNA were analyzed using previously tested and stored patient specimens, reference materials, and external quality controls. Results were compared with established reference assays used in accredited laboratories. Statistical analyses included positive, negative, and overall percent agreement (PPA, NPA, OPA), coefficients of variation (CV%), correlation and regression analyses and Bland-Altman bias estimation. For HIV-1 RNA, 19 of 20 positive and all 20 negative plasma samples were correctly identified by the ELITe MGB® assay, yielding a PPA of 95.0%, NPA of 100.0% and OPA of 97.5% (κ= 0.95). Intra-assay precision showed strong repeatability, with CVs of <1-3.9% for low-positive, 0.4-6.4% for medium-positive and <2% for high-positive specimens. Inter-assay reproducibility was consistent with CVs of 12.8% at low, 2.4% at medium, and 1.4% at high viral loads. Correlation analysis showed excellent concordance with the reference assay (p= 0.975, p< 0.001; R²= 0.95) and a mean bias of -0.40 log10 copies/mL in Bland-Altman analysis. Linearity was strong (R²= 0.97), confirming accurate quantification across the dynamic range with minor underestimation at higher dilutions. For HCV RNA, all 14 positive and 14 negative samples were correctly classified (PPA, NPA, and OPA= 100%; κ= 1.00). Intra-assay precision was excellent, with CVs around 2% for both low- and medium-positive samples, confirming consistent repeatability within a single run. Inter-assay reproducibility was equally robust, with CVs of 0.4-3.3% for low positives, 1.0-2.5% for medium and <1.1% for high-titer specimens. Correlation with the comparator method was strong (r= 0.956, p< 0.001; R²= 0.91) with a mean bias of -0.38 log10 IU/mL. Linearity analysis confirmed high proportionality between expected and measured concentrations (R²= 0.96). Deviations were negligible at low titers and slightly elevated at high loads but remained within acceptable limits. The HIV-1 and HCV ELITe MGB® assays on the BeGenius® platform demonstrated high accuracy, reproducibility and linearity, showing excellent correlation with reference methods. These results confirm that the ELITe BeGenius® system provides reliable and clinically valid viral-load measurements suitable for routine diagnostic use. Comprehensive laboratory verification of molecular assays under real-world conditions is crucial to ensure consistent performance, cross-platform comparability and reliable viral-load monitoring.