Direct blotting electrophoresis for sequencing and genotyping hepatitis C virus

Castelain, Sandrine; Khorsi, Hafida; Zawadzki, Patricia; Sueur, Jean-Marie; Capron, Jean‐Pierre; Eb, F.; Duverlie, Gilles

doi:10.1016/s0166-0934(97)02184-8

Cited by 7 publications

(3 citation statements)

References 18 publications

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“…The presence of HCV antibodies was determined and confirmed using two third-generation HCV EIA assays (Axsym ® HCV Version 3.0, Abbott, Wiesbaden, Germany and Vitros ® Anti-HCV reagent pack, Ortho-Clinical Diagnostic, High Wycombe, United Kingdom). HCV RNA was determined with a qualitative commercial assay (Cobas Amplicor HCV test Version 2.0, Roche Diagnostics, Meylan, France) and HCV genotyping was performed by direct sequencing, as described elsewhere [34]. The genotypes were distributed as follows: 11, 11, 11, 12, 10 and 2 samples of types 1a, 1b, 2, 3, 4 and 5, respectively.…”

Section: Patients and Clinical Samplesmentioning

confidence: 99%

A focus reduction neutralization assay for hepatitis C virus neutralizing antibodies

Fournier

Duverlie

François

et al. 2007

Virol J

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Background/Aim: The role of humoral immunity in hepatitis C virus (HCV) infection is poorly understood. Nevertheless, there is increasing interest in characterizing the neutralizing antibodies in the serum of HCV-infected patients. Focus reduction assays have been widely used to evaluate neutralizing antibody responses against a range of non-cytopathic viruses. Based on the recent development of a HCV cell culture system using the genotype 2 JFH-1-strain, we developed a focus reduction assay for HCV-neutralizing antibodies.

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Section: Patients and Clinical Samplesmentioning

confidence: 99%

A focus reduction neutralization assay for hepatitis C virus neutralizing antibodies

Fournier

Duverlie

François

et al. 2007

Virol J

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“…All patients chronically infected with HCV type 1 who were previously interferon-a2 (IFN-a2) non-responders (NR) were randomized and received either IFN-a2a (3 MU daily for 12 weeks, then three times per week), plus ribavirin (1,000-1,200 mg/day) (group B, bitherapy, n ¼ 15), or the same treatment plus amantadine hydrochloride (200 mg/day) (group T, tritherapy, n ¼ 12) for 48 weeks. Genotyping was carried out as previously described [Castelain et al, 1997].…”

Section: Patients and Samplesmentioning

confidence: 99%

Hepatitis C Virus p7 membrane protein quasispecies variability in chronically infected patients treated with interferon and ribavirin, with or without amantadine

Castelain¹,

Bonte²,

Pénin³

et al. 2006

Journal of Medical Virology

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A clinical study was carried out to compare the response rate of two groups of non-responder (NR) hepatitis C virus (HCV) genotype 1 chronically infected patients treated with interferon and ribavirin, with or without amantadine. The viral load decreased more markedly in the group treated by tritherapy including amantadine, but the response rate at the end of treatment was not significantly different between bitherapy and tritherapy. As amantadine could have an antiviral effect on the ion channel activity of the p7 HCV protein, the p7 quasispecies was characterized by cloning and sequencing. Sequence data were analyzed to determine the pattern and significance of p7 genetic heterogeneity and a possible relationship with therapy. Subtype differences were confirmed between p7 HCV genotypes 1a and 1b, and quasispecies analysis showed a reduction of genetic diversity in subtype 1a, but not 1b, during tritherapy. However, the absence of changes at numerous positions, as well as the conservative changes at other positions, indicated the high conservation of the p7 structure. Residue His-17, proposed to interact with amantadine, was fully conserved in both subtypes 1a and 1b, independently of amantadine administration. In conclusion, although the analysis of the p7 sequences revealed a selective pressure during therapy, no specific residues appeared to be linked to the effect of amantadine on viral decline. These results suggest that the potential antiviral effect of amantadine might be non-specific and related to a reduction in endosomal acidification and therefore reduced viral entry of HCV via its pH-dependent pathway.

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“…Genotyping was done by direct sequencing as described elsewhere (Castelain et al, 1997). Then, we compared our results with those obtained by the commercial HCV RNA quantitation assays described before.…”

Section: Hcv Genotype Reactivitymentioning

confidence: 99%

TaqMan amplification system with an internal positive control for HCV RNA quantitation

Castelain

Descamps

Thibault

et al. 2004

Journal of Clinical Virology

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Direct blotting electrophoresis for sequencing and genotyping hepatitis C virus

Cited by 7 publications

References 18 publications

A focus reduction neutralization assay for hepatitis C virus neutralizing antibodies

A focus reduction neutralization assay for hepatitis C virus neutralizing antibodies

Hepatitis C Virus p7 membrane protein quasispecies variability in chronically infected patients treated with interferon and ribavirin, with or without amantadine

TaqMan amplification system with an internal positive control for HCV RNA quantitation

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