Paul-Ehrlich-Institut, Section of Molecular Virology, Langen, Germany.
J Clin Virol. 2012 Dec;55(4):303-9. doi: 10.1016/j.jcv.2012.08.013. Epub 2012 Sep 12.
WHO International Standards (IS) are provided for the calibration and validation of diagnostic and screening assays, e.g. for hepatitis B virus (HBV). HBV forms numerous subgenotypes and the current IS for HBV DNA reflects subgenotype A2.
A reference panel with the most prevalent subgenotypes should facilitate evaluation of genotype-specific detection efficiencies.
215 HBV positive plasma samples collected worldwide were characterized for HBV markers and sequenced. Fifteen subgenotype A1, A2, B2, B4, C2, D1, D3, E, F2 and G samples were selected for the panel. The lyophilized samples were tested in parallel with the IS in an international collaborative study with 16 laboratories using 13 different nucleic acid amplification techniques (NATs).
Eight of 13 NAT had a HBV DNA detection efficiency which was independent of the genotype and consistent with the IS, while with five assays, certain deviations were noted, particularly with genotype F which was under quantitated or even missed by three assays. The panel was accepted by the WHO as the "1st WHO International Reference Panel for HBV Genotypes for HBV NAT-Based Assays".
The evaluation of HBV DNA assays should include many different genotypes. The WHO Reference Panel is universally available for manufacturers of HBV DNA assays, diagnostic laboratories and control authorities to facilitate standardized validation of HBV genotype specific detection efficiency of both diagnostic (quantitative and qualitative) and screening NAT assays.
世界卫生组织（WHO）国际标准（IS）用于校准和验证诊断和筛查检测，例如乙型肝炎病毒（HBV）。HBV 有许多亚型，目前用于 HBV DNA 的 IS 反映了 A2 亚型。
具有最常见亚型的参考面板应有助于评估基因型特异性检测效率。
对全球收集的 215 份 HBV 阳性血浆样本进行 HBV 标志物特征分析和测序。选择 15 份 A1、A2、B2、B4、C2、D1、D3、E、F2 和 G 亚型的样本作为该面板的一部分。将冻干样本与国际标准一起在国际合作研究中与 16 个实验室使用 13 种不同的核酸扩增技术（NAT）进行平行检测。
13 种 NAT 中有 8 种具有与基因型无关且与 IS 一致的 HBV DNA 检测效率，而对于 5 种检测方法，特别是对于基因型 F，检测结果存在一些偏差，这 5 种检测方法要么无法定量检测到，要么甚至漏检了 3 种检测方法。该面板被世卫组织接受为“用于基于 NAT 的 HBV DNA 检测的 1 型乙型肝炎病毒基因型 WHO 国际参考面板”。
HBV DNA 检测方法的评估应包括许多不同的基因型。世卫组织参考面板可供 HBV DNA 检测制造商、诊断实验室和控制机构使用，以促进诊断（定量和定性）和筛查 NAT 检测的 HBV 基因型特异性检测效率的标准化验证。
