Medizinische Universität Graz - Research portal
Selected Publication:
Stelzl, E.
Evaluation of two new assays for hepatitis C virus geno- and subtyping
[ Dissertation ] Graz Medical University; 2007.
- Authors Med Uni Graz:
- Advisor:
- Kessler Harald
- Reinthaler Franz
- Altmetrics:
- Abstract:
- It is estimated that more than 170 million people are infected with hepatitis C virus /HCV) worldwide. Treatment with interferon-alpha plus ribavirin is currently the standard therapy. The HCV genotype has been considered as major viral factor for tailoring Anti-HCV therapy. Furthermore, the HCV genotype and subtype are relevant to epidemiological studies, clinical management, pathogeneicity, and vaccine development. In this work, three different assays for determination of the HCV genotype and subtype, the routinely used TRUGENE HCV5NC Genotype Kit, the new VERSANT HCV Genotype 2.0 Assay (LiPA), and a new laboratory-development HCV BS5b sequencing assay designed for sequencing of the HCV NS5b region were investigated. A molecular diagnostics HCV genotyping proficiency program panel was used to determine accuracy. A total of 441 clinical samples obtained from patients with chronic HCV infection were collected at the molecular diagnostics laboratory, Institute of Hygiene, and tested for their HCV genotype and subtype with the routinely used TRUGENE HCV 5NC genotyping Kit. Samples for which no subtype was determined were retested using both of the new methods. All amplified samples from among the members of an HCV genotyping proficiency program panel that contained a single HCV genotype were subtyped correctly using all three HCV genotyping assays. With the TRUGENE HCV 5NC Genotyping Kit, the HCV subtype was determined in 357 of 441 of routine clinical samples. When the 84 samples with only genotype results were retested with the VERSANT HCV Genotype 2.0 Assay (LiPA), 61 could be further subtyped accurately. With the new laboratory-developed HCV NS5b sequencing assay, all of the 84 samples investigated could subtyped accurately. The two new methods advantages over the routinely used TRUGENE HCV 5NC Genotyping Kit in terms of genotyping and subtyping accuracy by utilizing part of the HV core region and NS5b region, respectively.