Reliable quantification of plasma HDV RNA is of paramount importance for treatment monitoring: A European multicenter study

Evelyn Stelzl; Sandra Ciesek; Markus Cornberg; Benjamin Maasoumy; Albert Heim; Michael Chudy; Antonella Olivero; Fabienne N Miklau; Annina Nickel; André Reinhardt; Michael Dietzsch; Harald H Kessler

doi:10.1016/j.jcv.2021.104932

Reliable quantification of plasma HDV RNA is of paramount importance for treatment monitoring: A European multicenter study

J Clin Virol. 2021 Sep:142:104932. doi: 10.1016/j.jcv.2021.104932. Epub 2021 Jul 24.

Authors

Affiliations

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.

PMID: 34333392
DOI: 10.1016/j.jcv.2021.104932

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.

Keywords: 1st WHO International Standard for HDV RNA; Extraction platforms; Hepatitis D virus (HDV); IU/mL; Quantitative real-time pcr.

Publication types

Multicenter Study
Research Support, Non-U.S. Gov't

MeSH terms

Hepatitis D* / diagnosis
Hepatitis D* / drug therapy
Hepatitis Delta Virus / genetics
Humans
Pharmaceutical Preparations*
RNA, Viral
Reproducibility of Results
Viral Load

Substances

Pharmaceutical Preparations
RNA, Viral