Evelyn Stelzl1, Sandra Ciesek2, Markus Cornberg3, Benjamin Maasoumy3, Albert Heim4, Michael Chudy5, Antonella Olivero6, Fabienne N Miklau1, Annina Nickel7, André Reinhardt7, Michael Dietzsch7, Harald H Kessler8. 1. Research Unit Molecular Diagnostics, Diagnostic and Research Center for Molecular Biomedicine, Medical University of Graz, Austria. 2. Institute of Virology, University Hospital Essen, Germany (currently Institute for Medical Virology, University Hospital Frankfurt, Germany). 3. Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Germany. 4. Department for Virology, Hannover Medical School, Germany. 5. Section of Molecular Virology, Paul-Ehrlich-Institut, Langen, Germany. 6. University of Torino, Department of Medical Sciences, Laboratory of Molecular Hepatology and Gastroenterology, Torino, Italy. 7. Roboscreen GmbH, Leipzig, Germany. 8. Research Unit Molecular Diagnostics, Diagnostic and Research Center for Molecular Biomedicine, Medical University of Graz, Austria. Electronic address: harald.kessler@medunigraz.at.
Abstract
OBJECTIVES: Quantification of plasma hepatitis D virus (HDV) RNA is the essential tool for patient management under antiviral therapy. The aim of this European multicenter study was to improve the comparability of quantitative results reported by different laboratories using the CE/IVD-labeled RoboGene HDV RNA Quantification Kit 2.0 (Roboscreen GmbH) with different manual or automated nucleic acid extraction protocols/platforms and amplification/detection devices. METHODS: For harmonization of HDV RNA concentrations obtained by different protocols, correction factors (CF) were determined using the 1st WHO International Standard for HDV RNA. The limit of detection (LOD) and accuracy were determined for each protocol by using reference material. Furthermore, clinical samples were analyzed and results compared. RESULTS: The CF ranged from 20 to 1,870 depending on the protocol used. The LOD was found between 4 and 450 IU/ml. When accuracy was tested, external quality control (EQC) samples containing low HDV RNA concentrations were not detected by those protocols with higher LODs. For EQC samples, the maximum standard deviation of HDV RNA concentrations was found to be 0.53 log10 IU/ml, for clinical samples 0.87 log10 IU/mL. CONCLUSION: To ensure reliability in quantification of HDV RNA, any modification of the extraction and amplification/detection protocol validated by the manufacturer requires revalidation. With the 1st WHO International Standard for HDV RNA, the CF could easily be calculated leading to harmonization of quantitative results. This warrants both accurate monitoring of response to existing anti-HDV treatment and comparability of study results investigating novel anti-HDV drugs.
OBJECTIVES: Quantification of plasma hepatitis D virus (HDV) RNA is the essential tool for patient management under antiviral therapy. The aim of this European multicenter study was to improve the comparability of quantitative results reported by different laboratories using the CE/IVD-labeled RoboGene HDV RNA Quantification Kit 2.0 (Roboscreen GmbH) with different manual or automated nucleic acid extraction protocols/platforms and amplification/detection devices. METHODS: For harmonization of HDV RNA concentrations obtained by different protocols, correction factors (CF) were determined using the 1st WHO International Standard for HDV RNA. The limit of detection (LOD) and accuracy were determined for each protocol by using reference material. Furthermore, clinical samples were analyzed and results compared. RESULTS: The CF ranged from 20 to 1,870 depending on the protocol used. The LOD was found between 4 and 450 IU/ml. When accuracy was tested, external quality control (EQC) samples containing low HDV RNA concentrations were not detected by those protocols with higher LODs. For EQC samples, the maximum standard deviation of HDV RNA concentrations was found to be 0.53 log10 IU/ml, for clinical samples 0.87 log10 IU/mL. CONCLUSION: To ensure reliability in quantification of HDV RNA, any modification of the extraction and amplification/detection protocol validated by the manufacturer requires revalidation. With the 1st WHO International Standard for HDV RNA, the CF could easily be calculated leading to harmonization of quantitative results. This warrants both accurate monitoring of response to existing anti-HDV treatment and comparability of study results investigating novel anti-HDV drugs.
Authors: Antonella Olivero; Chiara Rosso; Alessia Ciancio; Maria Lorena Abate; Aurora Nicolosi; Giulia Troshina; Angelo Armandi; Davide Giuseppe Ribaldone; Giorgio Maria Saracco; Elisabetta Bugianesi; Mario Rizzetto; Gian Paolo Caviglia Journal: Biomedicines Date: 2022-03-29
Authors: Carla Usai; Upkar S Gill; Anna C Riddell; Tarik Asselah; Patrick T Kennedy Journal: Aliment Pharmacol Ther Date: 2022-03-16 Impact factor: 9.524