Research Unit Molecular Diagnostics, Diagnostic and Research Center for Molecular Biomedicine, Medical University of Graz, Austria.
Institute of Virology, University Hospital Essen, Germany (currently Institute for Medical Virology, University Hospital Frankfurt, Germany).
J Clin Virol. 2021 Sep;142:104932. doi: 10.1016/j.jcv.2021.104932. Epub 2021 Jul 24.
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.
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.
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 log IU/ml, for clinical samples 0.87 log IU/mL.
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.
检测血浆中丁型肝炎病毒（HDV）RNA 的含量是抗病毒治疗患者管理的重要手段。本欧洲多中心研究旨在通过使用经 CE/IVD 认证的 RoboGene HDV RNA 定量试剂盒 2.0（Roboscreen GmbH），并结合不同的手动或自动化核酸提取方案/平台和扩增/检测设备，提高不同实验室报告的定量结果的可比性。
为了协调不同方案获得的 HDV RNA 浓度，使用第 1 个世界卫生组织（WHO）HDV RNA 国际标准确定校正因子（CF）。通过使用参考物质，确定了每种方案的检测限（LOD）和准确性。此外，还分析了临床样本并进行了结果比较。
CF 值因使用的方案而异，范围从 20 到 1870。LOD 范围为 4 至 450 IU/ml。当测试准确性时，那些 LOD 较高的方案无法检测到含有低 HDV RNA 浓度的外部质量控制（EQC）样本。对于 EQC 样本，HDV RNA 浓度的最大标准偏差为 0.53 log IU/ml，对于临床样本为 0.87 log IU/mL。
为了确保 HDV RNA 定量的可靠性，任何经制造商验证的提取和扩增/检测方案的修改都需要重新验证。通过第 1 个 WHO 国际 HDV RNA 标准，可以轻松计算 CF，从而实现定量结果的协调。这不仅保证了对现有抗 HDV 治疗反应的准确监测，还保证了正在研究新型抗 HDV 药物的研究结果的可比性。
