Introduction: A reliable quantification of hepatitis D virus (HDV) RNA is of paramount importance for monitoring patients under antiviral therapy. This quality control study compares the diagnostic performances of quantitative HDV-RNA assays used in clinical practice. Methods: Two HDV-RNA sample panels were quantified in 30 centers by RoboGene (N = 9 laboratories), EurobioPlex (N = 7), RealStar (N = 4), AltoStar (N = 1), Bosphore (N = 3), Bosphore-on-InGenius (N = 1), Dia.Pro (N = 2), Nuclear-Laser-Medicine (N = 1) and 3 in-house assays. Panel A and B comprised 8 serial dilutions of WHO/HDV standard (range: 0.5–5.0 log10 IU/ml) and 20 clinical samples (range: 0.5–6.0 log10 IU/ml), respectively. The following parameters were determined: sensitivity by 95 % LOD (limit of detection), precision by intra- and inter-run CV (coefficient of variation), accuracy by the differences between expected-observed HDV-RNA, linearity by linear regression analysis. Results: 95 % LOD varied across assays and centers underlining heterogeneous sensitivities: AltoStar had the lowest 95 % LOD (3 IU/ml) followed by RealStar (10 [min–max: 3–316] IU/ml), Bosphore-on-InGenius (10 IU/ml), RoboGene (31 [3–316] IU/ml), Nuclear-Laser-Medicine (31 IU/ml) and EuroBioplex (100 [100–316] IU/ml). Moreover, 6 assays (RoboGene, EurobioPlex, RealStar, AltoStar, Nuclear-Laser-Medicine and In-house) showed <0.5 log10 IU/ml differences between expected and observed HDV-RNA for all dilutions while other assays had >1 log10 IU/ml underestimations. RealStar, Bosphore-on-InGenius and EurobioPlex had the highest precision (mean intra-run CV < 20 %). Inter-run CV was higher for all assays, with CVs < 25 % for RealStar, AltoStar, Nuclear-Laser-Medicine and EurobioPlex. Seven assays (RoboGene/AltoStar/RealStar/EurobioPlex/Nuclear-Laser-Medicine/In-house) showed a good linearity (R2 > 0.90), but for HDV-RNA < 1000 IU/ml only Bosphore-on-InGenius, AltoStar, RealStar and Robogene showed a R2 > 0.85. Conclusions: This study underlines heterogeneous sensitivities (inter- and intraassays), that could hamper proper HDV-RNA quantification, particularly at low viral loads. This raises the need to improve the diagnostic performance of most assays for properly identifying virological response to anti-HDV drugs.
Comparison of diagnostic performances of HDV-RNA quantification assays used in clinical practice: Results from a national quality control multicenter study / Salpini, R.; Piermatteo, L.; Caviglia, G. P.; Bertoli, A.; Brunetto, M. R.; Bruzzone, B.; Callegaro, A.; Caudai, C.; Cavallone, D.; Chessa, L.; Coghe, F.; Coppola, N.; Cuomo, N.; D'Anna, S.; Di Stefano, M.; Facchetti, F.; Farina, C.; Ferraro, D.; Franchin, E.; Francisci, D.; Galli, S.; Garbuglia, A. R.; Gennari, W.; Ghisetti, V.; Lampertico, P.; Lo Caputo, S.; Marascio, N.; Menzo, S.; Micheli, V.; Niro, G. A.; Olivero, A.; Paba, P.; Palermo, C. I.; Palmieri, O.; Paolucci, S.; Pisaturo, M.; Pollicino, T.; Raffa, G.; Santantonio, T.; Torre, G.; Turriziani, O.; Uzzau, S.; Uceda Renteria, S. C.; Vatteroni, M.; Zazzi, M.; Craxi, A.; Ceccherini-Silberstein, F.; Svicher, V.; Arosio, M.; Bastianelli, S.; Gentile, A.; Giardina, F. A. M.; Gidari, A.; Govoni, R.; Ibba, G.; Loglio, A.; Lombardi, A.; Mascarella, C.; Maggi, F.; Matera, G.; Mazzei, C.; Milia, M. G.; Quirino, A.; Raddi, A.; Scioscia, R.; Tagliazucchi, S.; Totaro, M.; Valaperta, R.. - In: JOURNAL OF CLINICAL VIROLOGY. - ISSN 1386-6532. - 180:(2025). [10.1016/j.jcv.2025.105850]
Comparison of diagnostic performances of HDV-RNA quantification assays used in clinical practice: Results from a national quality control multicenter study
Uzzau S.;Ibba G.;
2025-01-01
Abstract
Introduction: A reliable quantification of hepatitis D virus (HDV) RNA is of paramount importance for monitoring patients under antiviral therapy. This quality control study compares the diagnostic performances of quantitative HDV-RNA assays used in clinical practice. Methods: Two HDV-RNA sample panels were quantified in 30 centers by RoboGene (N = 9 laboratories), EurobioPlex (N = 7), RealStar (N = 4), AltoStar (N = 1), Bosphore (N = 3), Bosphore-on-InGenius (N = 1), Dia.Pro (N = 2), Nuclear-Laser-Medicine (N = 1) and 3 in-house assays. Panel A and B comprised 8 serial dilutions of WHO/HDV standard (range: 0.5–5.0 log10 IU/ml) and 20 clinical samples (range: 0.5–6.0 log10 IU/ml), respectively. The following parameters were determined: sensitivity by 95 % LOD (limit of detection), precision by intra- and inter-run CV (coefficient of variation), accuracy by the differences between expected-observed HDV-RNA, linearity by linear regression analysis. Results: 95 % LOD varied across assays and centers underlining heterogeneous sensitivities: AltoStar had the lowest 95 % LOD (3 IU/ml) followed by RealStar (10 [min–max: 3–316] IU/ml), Bosphore-on-InGenius (10 IU/ml), RoboGene (31 [3–316] IU/ml), Nuclear-Laser-Medicine (31 IU/ml) and EuroBioplex (100 [100–316] IU/ml). Moreover, 6 assays (RoboGene, EurobioPlex, RealStar, AltoStar, Nuclear-Laser-Medicine and In-house) showed <0.5 log10 IU/ml differences between expected and observed HDV-RNA for all dilutions while other assays had >1 log10 IU/ml underestimations. RealStar, Bosphore-on-InGenius and EurobioPlex had the highest precision (mean intra-run CV < 20 %). Inter-run CV was higher for all assays, with CVs < 25 % for RealStar, AltoStar, Nuclear-Laser-Medicine and EurobioPlex. Seven assays (RoboGene/AltoStar/RealStar/EurobioPlex/Nuclear-Laser-Medicine/In-house) showed a good linearity (R2 > 0.90), but for HDV-RNA < 1000 IU/ml only Bosphore-on-InGenius, AltoStar, RealStar and Robogene showed a R2 > 0.85. Conclusions: This study underlines heterogeneous sensitivities (inter- and intraassays), that could hamper proper HDV-RNA quantification, particularly at low viral loads. This raises the need to improve the diagnostic performance of most assays for properly identifying virological response to anti-HDV drugs.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
