Alberta Provincial Laboratory for Public Health, Edmonton, Alberta, Canada.
Alberta Provincial Laboratory for Public Health, Calgary, Alberta, Canada.
J Clin Virol. 2014 Jul;60(3):301-4. doi: 10.1016/j.jcv.2014.04.005. Epub 2014 Apr 14.
Hepatitis C (HCV) genotyping is important for treatment planning. The Abbott m2000 RealTime HCV Genotype II assay is a PCR-based assay targeting specific regions of the 5'NCR gene for genotypes 1-6, and the NS5b gene for subgenotypes 1a/1b. However, not all genotypes can be resolved, with results being reported as: 'indeterminate', 'mixed', 'genotype X reactivity with Y', or just the major genotype 1 alone.
To assess the supplementary testing required for these unresolved HCV genotypes, these samples were tested further using an in-house core/E1 sequencing assay. The resulting genotypes/subgenotypes were assigned using phylogenetic analysis with reference HCV genotype sequences. Additional testing was conducted using the INNO-LiPA HCV II assay for truly mixed genotypes.
Out of 1052 samples tested, 89 (8.5%) underwent further sequencing to determine the HCV genotype: 16 that were 'indeterminate' on the m2000, were mostly genotype 2s and 3s by sequencing; 12 that were 'mixed', were mostly one of the genotypes reported in the mixture; 7 that were 'X reactivity with Y', were usually genotype X; 54 that gave just a major genotype 1 result were mostly 1a, with some 6 and 1b, and a few 1c. For three truly mixed genotypes, additional testing using the VERSANT(®) HCV Genotype Assay (LiPA) 2.0, showed two mixed 1 and 3, and one indistinguishable 6c-6l genotypes.
The Abbott m2000 RealTime HCV Genotype II assay can resolve most (∼90%) HCV genotypes. However in 9-10% of cases, to fully resolve the genotype, additional testing is required.
丙型肝炎（HCV）基因分型对于治疗方案的制定很重要。Abbott m2000 RealTime HCV Genotype II 检测是一种基于 PCR 的检测方法，针对 5'NCR 基因的特定区域检测基因型 1-6，以及 NS5b 基因的亚基因型 1a/1b。然而，并非所有基因型都能得到确定，结果报告为“不确定”、“混合”、“基因型 X 与 Y 的反应”或仅为主要基因型 1 单独存在。
为了评估这些未解决的 HCV 基因型所需的补充检测，这些样本进一步使用内部核心/E1 测序检测进行了检测。使用与参考 HCV 基因型序列的系统发育分析来分配产生的基因型/亚基因型。使用 INNO-LiPA HCV II 检测对真正的混合基因型进行了额外的检测。
在检测的 1052 个样本中，有 89 个（8.5%）进行了进一步的测序以确定 HCV 基因型：在 m2000 上显示为“不确定”的 16 个，通过测序主要为基因型 2 和 3；12 个“混合”，主要是混合物中报告的基因型之一；7 个“X 与 Y 的反应”，通常是基因型 X；54 个仅显示主要基因型 1 的结果，主要是 1a，有一些 6 和 1b，还有一些 1c。对于三个真正的混合基因型，使用 VERSANT®HCV Genotype Assay (LiPA) 2.0 进行的额外检测显示，两种混合 1 和 3，一种无法区分的 6c-6l 基因型。
Abbott m2000 RealTime HCV Genotype II 检测可以确定大多数（约 90%）HCV 基因型。然而，在 9-10%的情况下，需要进行额外的检测才能完全确定基因型。
