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
Articles in the same Issue
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Articles in the same Issue
- Associations between folate, vitamin B12, homocysteine and pathologies related to aging: the need to consider complex nutrient-nutrient and gene-nutrient interactions and the functional and socio-economic determinants in population-based studies
- Hyperhomocysteinemia and cardiovascular risk in postmenopausal women: the role of folate supplementation
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- Association of thyroid dysfunction with vitamin B12, folate and plasma homocysteine levels in the elderly: a population-based study in Sicily
- Desmond (Des) Kenny MSc MCB FICI FRCPath Eur Clin Chem, 1941–2006
- Antibodies to citrullinated peptides: a significant step forward in the early diagnosis of rheumatoid arthritis
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- Quantification of relative changes in specific mRNAs from frozen whole blood – methodological considerations and clinical implications
- HER-2 protein concentrations in breast cancer cells increase before immunohistochemical and fluorescence in situ hybridization analysis turn positive
- Distinct alleles of mannose-binding lectin (MBL) and surfactant proteins A (SP-A) in patients with chronic cavitary pulmonary aspergillosis and allergic bronchopulmonary aspergillosis
- Novel molecular defect in the platelet ADP receptor P2Y12 of a patient with haemorrhagic diathesis
- Retrospective study of monoclonal gammopathies detected in the clinical laboratory of a Spanish healthcare district: 14-year series
- Renal impairment compromises the use of total homocysteine and methylmalonic acid but not total vitamin B12 and holotranscobalamin in screening for vitamin B12 deficiency in the aged
- Diagnostic value of calculated testosterone indices in the assessment of polycystic ovary syndrome
- Amniotic fibronectin fragmentation and expression of its domains, sialyl and fucosyl glycotopes associated with pregnancy complicated by intrauterine infection
- Predicted influence of sample hematocrit on injected mass of internal standard in mass spectrometry assays utilizing simple protein precipitation for sample preparation
- Evaluation of cardiovascular risk factors and related clinical markers in healthy young Japanese adults
- Cardiac natriuretic peptides after myocardial infarction: relationship with infarct size, left ventricular function and remodelling assessed by 99mTc-sestamibi gated-single photon emission tomography
- Plasma homocysteine cutoff values for venous thrombosis
- Lipid transfer protein activities in subjects with impaired glucose tolerance
- Heat shock protein 70-1 gene expression in pediatric heart surgery using blood cardioplegia
- Biochemical reference intervals for sex hormones with a new AutoDelfia method in aged men
- The Italian External Quality Control Programme for cystic fibrosis molecular diagnosis: 4 years of activity
- Standardization of ischemia-modified albumin testing: adjustment for serum albumin
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- To the Editor: Author reply; Nakano et al. Clin Chem Lab Med 2006;44(5):522–532
- Accreditation of medical laboratories in the European Union
- Prevalence-dependent decision limits for the early detection of type 2 diabetes mellitus in venous blood, venous plasma and capillary blood during glucose challenge
