Determination of the hepatitis C virus subtype: comparison of sequencing and reverse hybridization assays
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Evelyn Stelzl
Abstract
Background: Hepatitis C virus (HCV) genotyping and accurate subtyping is becoming increasingly relevant to epidemiological studies, clinical management, pathogenicity, and vaccine development.
Methods: The TRUGENE® HCV 5′NC Genotyping Kit, the new VERSANT® HCV Genotype 2.0 Assay (LiPA), and a new laboratory-developed HCV NS5b sequencing assay designed for automated sequencing of the HCV NS5b region were used. Clinical samples and a molecular diagnostics HCV genotyping proficiency program panel were used to determine accuracy and differentiate performance characteristics of the three methods.
Results: All amplified samples from among the members of a HCV genotyping proficiency program panel that contained a single HCV genotype were subtyped correctly using all three HCV genotyping assays. With the TRUGENE® HCV 5′NC 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 84 could be subtyped accurately.
Conclusions: The two new methods show advantages over the routinely used TRUGENE® HCV 5′NC Genotyping Kit in terms of genotyping and subtyping accuracy by utilizing part of the HCV core region and NS5b region, respectively.
Clin Chem Lab Med 2007;45:167–70.
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©2007 by Walter de Gruyter Berlin New York
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