HCV

Hepatitis C virus

Hepatitis C (HCV) virus infection is mostly chronic and can lead to the development of liver cirrhosis and hepatocellular carcinoma. The virus is highly heterogenous, and at least 6 major genotypes have been recognized, each with a large number of subtypes. The likelihood of a response to antiviral therapy is clearly associated with the HCV genotype, and genotyping is clinically important in choosing the appropriate HCV therapy. At present no effective vaccine is available, but multiple highly effective directly-acting antiviral agents have recently been developed.

DDL is an expert laboratory for the analysis of resistance-associated mutations in HCV and provides its support to many HCV DAA studies, involving numerous NS3, NS5A and NS5B inhibitors from more than 10 different sponsors.

Genotyping

  • Simultaneous identification of genotypes 1, 2, 3, 4, 5, and 6  including all relevant subtypes
  • Based on a pan-genotypic RT-PCR followed by sequence analysis of partial NS5B region (genotyping reference region)
  • Often used as confirmation assay to confirm HCV genotyping results as determined by standard HCV genotyping assays, such as Versant HCV LiPA or Abbott HCV genotyping.

Screening for mutations associated with resistance to antiviral therapy

  • Based on RT-PCR amplification of the relevant region of the genome followed by Sanger or deep sequencing (Illumina MiSeq).
  • NS3, NS4A, NS4B, NS5A and NS5B regions can be analysed in virtually all major genotypes
Genomic region
Genotype NS3-4a NS4B NS5A NS5B
1
2
3
4
5 In development
6 In development
  • Monitoring of antiviral therapy by population or deep sequencing
  • Tailor-made reporting in tailor-made (including FDA-EMA compatible) formats

Phylogenetic analysis

  • Determination of relatedness between different strains
  • Used to confirm that samples are derived from the same patient or from a common source of infection.
  • For outbreak management and epidemiologic studies
  • Based on RT PCR followed by DNA sequencing analysis
  • can be targeted on any viral genome region (NS5B is preferred target)

Samples

Serum or EDTA plasma. Heparinized blood can not be used.

Respiratory infections

DDL has extensive and expanding experience in molecular diagnosis and characterization of respiratory infections.

Broad viral screening can be performed using e.g., the Luminex xTAG RVP FAST v2 assay, detecting: RSVA/B, influenza A (H1, H1pdm2009, H3), influenza B, parainfluenza (1-4), metapneumovirus, adenovirus, entero-rhinovirus, coronavirus (NL63, HKU1, 229E, OC43), bocavirus.

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