Research Unit Molecular Diagnostics and Molecular Diagnostics Laboratory, IHMEM, Medical University of Graz, Graz, Austria.
J Clin Virol. 2013 Nov;58(3):522-7. doi: 10.1016/j.jcv.2013.09.005. Epub 2013 Sep 12.
Accurate quantitation of hepatitis C virus (HCV) RNA is mandatory for the management of anti-HCV therapy.
The genotype-dependent performance of the new commercially available VERSANT HCV RNA 1.0 Assay (kPCR) and the COBAS AmpliPrep/COBAS TaqMan HCV Quantitative Test, version 2.0 was investigated.
The molecular assays for quantitation of HCV RNA were performed according to the manufacturer's package insert instructions. HCV genotypes/subtypes/isolates, and mutations in the 5'NCR were detected by direct sequencing.
When members of a worldwide HCV performance panel including HCV subtypes 1a, 1b, 2a, 3b, and 4a were tested with the Siemens assay and the results were compared with those obtained by the Roche assay, the mean log10 unit differences for members containing HCV subtypes 1a, 1b, 3b, and 4a were found to be within ±0.5 log(10) units. For the panel member containing HCV subtype 2a, the HCV RNA concentration was found to be >0.5 log(10) units lower with the Siemens assay. When clinical samples were tested, the HCV RNA concentration of all samples containing HCV subtype 2a were found to be >0.5 log(10) units lower with the Siemens assay while that of certain HCV subtype 3a and 4a isolates were found to be >1.0 log(10) units lower.
The VERSANT HCV RNA 1.0 Assay substantially underestimates HCV RNA concentrations in HCV subtype 2a samples and in HCV subtype 3a and 4a samples containing certain isolates. This may be caused by mismatches with the target sequences due to the primer and/or probe design.
准确定量丙型肝炎病毒 (HCV) RNA 是管理抗 HCV 治疗的必要条件。
研究新的商业化 VERSANT HCV RNA 1.0 检测 (kPCR) 和 COBAS AmpliPrep/COBAS TaqMan HCV 定量检测，版本 2.0 在依赖基因型方面的性能。
根据制造商的说明书进行 HCV RNA 定量的分子检测。通过直接测序检测 HCV 基因型/亚型/分离株和 5'NCR 突变。
当用西门子检测对包括 HCV 亚型 1a、1b、2a、3b 和 4a 的全球 HCV 性能面板成员进行检测，并将结果与罗氏检测的结果进行比较时，发现包含 HCV 亚型 1a、1b、3b 和 4a 的成员的平均对数 10 个单位差异在 ±0.5 log(10) 个单位内。对于包含 HCV 亚型 2a 的面板成员，西门子检测发现 HCV RNA 浓度低 >0.5 log(10) 个单位。当检测临床样本时，发现所有包含 HCV 亚型 2a 的样本的 HCV RNA 浓度均低 >0.5 log(10) 个单位，而某些 HCV 亚型 3a 和 4a 分离株的浓度则低 >1.0 log(10) 个单位。
VERSANT HCV RNA 1.0 检测严重低估了 HCV 亚型 2a 样本和包含某些分离株的 HCV 亚型 3a 和 4a 样本中的 HCV RNA 浓度。这可能是由于引物和/或探针设计与靶序列不匹配所致。
