Laboratory of Virology, Institute for Biology and Pathology, Grenoble Alpes University Hospital, Grenoble, France.
Department of Hepatology, Grenoble Alpes University Hospital, Grenoble, France.
PLoS One. 2018 Mar 22;13(3):e0194396. doi: 10.1371/journal.pone.0194396. eCollection 2018.
Direct-acting antiviral (DAA) drug performances depend on the viral genotype. So international recommendations give typing of the virus a prerequisite for treatment choice and patient management. Commercially available HCV genotyping kits are scarce and this analysis is often in-house using tedious PCRs and Sanger sequencing, leading to a lack of standardization. A newly commercialized HCV genotyping assay based on real-time PCR has been developed by Roche Diagnostics (Mannheim, Germany). We compared this new assay with our in-house PCRs -sequencing technique on 101 regular samples and 81 LiPA failures or low viral load samples. No genotype or 1a/1b subtype mismatch was observed. Two samples were misidentified at the subtype level without clinical impact. Three genotype 1b and two genotype 1a samples with low viral load could not be subtyped. Nevertheless, 13 (13%) samples from the regular panel and 35 (43%) from the more difficult-to-type panels failed to give results on first pass with the Roche kit. Failures were mostly associated with genotype 3 subtype a, with genotype 4 subtype non-a, or with viral loads <200 IU/mL (p = 0.0061). The workflow allowed a non-specialized technician to obtain results in less than 4 hours whereas 2 to 3 days and experienced staff were required with the in-house assay. In conclusion, the Roche cobas® HCV GT kit is easy and rapid to use and provides reliable results. The high rate of uninterpretable results particularly for low viral load samples and less frequent genotypes, and the absence of subtyping for non-genotype 1 could require sending complex samples to a specialized laboratory.
直接作用抗病毒 (DAA) 药物的疗效取决于病毒基因型。因此，国际指南规定在选择治疗方案和管理患者时，应先对病毒进行分型。市售的 HCV 基因分型试剂盒稀缺，而这种分析通常在内部使用繁琐的 PCR 和 Sanger 测序进行，导致缺乏标准化。罗氏诊断公司（德国曼海姆）开发了一种新的基于实时 PCR 的 HCV 基因分型检测方法。我们将该新检测方法与我们内部的 PCR-测序技术在 101 个常规样本和 81 个 LiPA 失败或低病毒载量样本中进行了比较。未观察到基因型或 1a/1b 亚型不匹配。有 2 个样本在亚类水平上被错误鉴定，但无临床影响。3 个低病毒载量的 1b 基因型和 2 个 1a 基因型样本无法进行亚型分型。然而，常规样本组中有 13（13%）个样本和更难分型的样本组中有 35（43%）个样本在初次检测时罗氏试剂盒无法给出结果。失败主要与基因型 3 亚型 a、基因型 4 非-a 亚型或病毒载量 <200 IU/mL（p = 0.0061）有关。该工作流程允许非专业技术人员在不到 4 小时内获得结果，而使用内部检测方法则需要 2 至 3 天和经验丰富的工作人员。总之，罗氏 cobas® HCV GT 试剂盒易于使用且快速，可提供可靠的结果。对于低病毒载量样本和不太常见的基因型，无法解释结果的比例较高，并且不能对非 1 型进行亚型分型，这可能需要将复杂样本送到专门的实验室进行检测。
