Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Zurl, H.
Determination of the hepatitis C virus genotype/subtype: evaluation of a new assay based on primer-specific real-time polymerase chain reaction
Humanmedizin; [ Diplomarbeit ] Graz Medical University; 2016. pp.
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- Autor*innen der Med Uni Graz:
- Zurl Hanna
- Betreuer*innen:
- Kessler Harald
- Stelzl Evelyn
- Altmetrics:
- Abstract:
- Background: Anti-hepatitis C virus treatment has undergone major progress recently through introduction of effective direct-acting antivirals. The hepatitis C virus (HCV) genotype and HCV subtypes 1a and 1b must be determined prior to treatment initiation as it is still essential for the choice of therapy. Objectives: The aim of this study was to evaluate the performance of the new cobas® HCV GT (Roche Molecular Systems) assay and to compare it to alternative assays used in the routine diagnostic laboratory. Methods: Accuracy was tested using the Quality Control for Molecular Diagnostics (QCMD) HCV Genotyping Proficiency Program 2014 and 2015 panels. For clinical evaluation, 183 residual routine clinical samples were tested and results were compared to those obtained by the VERSANT® HCV Genotype 2.0 Assay (LiPA) (Siemens Healthcare Diagnostics) and the TRUGENE® HCV 5’NC Genotyping Kit (Siemens). All samples showing discrepant results were additionally investigated by home-brew NS5B sequencing. In addition, total times for completion including hands-on times were compared. Results: When accuracy was tested, the cobas® HCV GT assay reported correct results for panel members containing HCV genotype 3, HCV genotype 4, and HCV genotype 1 subtypes 1a and 1b but the assay could not detect low titer panel members containing HCV genotype 5 correctly. When the clinical performance of the cobas® HCV GT was investigated, 160 out of 183 clinical samples showed concordant results when compared with alternative assays. Seven samples showed indeterminate results with the cobas® HCV GT and 16 samples showed discordant results with at least one of the comparator assays. When times-to-results were compared, the cobas® HCV GT could be completed faster than the comparator assays but hands-on time was similar for all assays. Conclusions: The new cobas® HCV GT assay showed a good clinical performance and proved to be suitable for use in the routine diagnostic laboratory. The new assay allows for accurate determination of the HCV genotypes 2, 3, and 4 and it is able to discriminate HCV genotype 1 subtype 1a and 1b.