Japanese Reference Panel of Blood Specimens for Evaluation of Hepatitis C Virus RNA and Core Antigen Quantitative Assays

Murayama, Asako; Sugiyama, Nao; Watashi, Koichi; Masaki, Takahiro; Suzuki, Ritsuro; Aizaki, Hideki; Mizuochi, Toshiaki; Wakita, Takaji; Kato, Takanobu

doi:10.1128/jcm.00487-12

Cited by 38 publications

(36 citation statements)

References 34 publications

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“…the "grey-zone") were retested per manufacturer recommendations where sample was available. For participants with quantifiable HCV RNA, and yet consistently negative HCVcAg results, the HCV Core gene was amplified and sequenced using previously published methods where sample was available [15,17] to identify potential mutations in the antibody binding region of the ARCHITECT HCV Ag assay [18][19][20].…”

Section: Study Assessmentsmentioning

confidence: 99%

Hepatitis C Virus Core Antigen: A Simplified Treatment Monitoring Tool, including for Post-Treatment Relapse

Lamoury¹,

Soker²,

Martinez³

et al. 2016

Journal of Hepatology

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Background: Simple, affordable diagnostic tools are essential to facilitate global hepatitis C virus (HCV) elimination efforts. Objectives: This study evaluated the clinical performance of core antigen (HCVcAg) assay from plasma samples to monitor HCV treatment efficacy and HCV viral recurrence. Study design: Plasma samples from a study of response-guided pegylated-interferon/ribavirin therapy for people who inject drugs with chronic HCV genotype 2/3 infection were assessed for HCV RNA (AmpliPrep/COBAS Taqman assay, Roche) and HCVcAg (ARCHITECT HCV Ag, Abbott Diagnostics) during and after therapy. The sensitivity and specificity of the HCVcAg assay was compared to the HCV RNA assay (gold standard). Results: A total of 335 samples from 92 enrolled participants were assessed (mean 4 time-points per participant). At baseline, end of treatment response (ETR) and sustained virological response (SVR) visits, the sensitivity of the HCVcAg assay with quantifiable HCV RNA threshold was 94% (95% CI: 88%, 98%), 56% (21%, 86%) and 100%, respectively. The specificity was between 98 to 100% for all time-points assessed. HCVcAg accurately detected all six participants with viral recurrence, demonstrating 100% sensitivity and specificity. One participant with detectable (non-quantifiable) HCV RNA and non-reactive HCVcAg at SVR12 subsequently cleared HCV RNA at SVR24. Conclusions: HCVcAg demonstrated high sensitivity and specificity for detection of pre-treatment and posttreatment viraemia. This study indicates that confirmation of active HCV infection, including recurrent viraemia, by HCVcAg is possible. Reduced on-treatment sensitivity of HCVcAg may be a clinical advantage given the moves toward simplification of monitoring schedules.

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Section: Study Assessmentsmentioning

confidence: 99%

Hepatitis C Virus Core Antigen: A Simplified Treatment Monitoring Tool, including for Post-Treatment Relapse

Lamoury¹,

Soker²,

Martinez³

et al. 2016

Journal of Hepatology

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“…The concentration of HCV core antigen (Ag) in the culture media and in the cell lysate was measured using a chemiluminescent enzyme immunoassay (CLEIA; Lumipulse Ortho HCV antigen; Fujirebio, Tokyo, Japan) in accordance with the manufacturer's instructions (37).…”

Section: Methodsmentioning

confidence: 99%

Amino Acid Mutations in the NS4A Region of Hepatitis C Virus Contribute to Viral Replication and Infectious Virus Production

Murayama

Sugiyama

Suzuki

et al. 2017

J Virol

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Hepatitis C virus (HCV) strain JFH-1, which belongs to genotype 2a, replicates autonomously in cultured cells, whereas another genotype 2a strain, J6CF, does not. Previously, we found that replacement of the NS3 helicase and NS5B-to-3=X regions of J6CF with those of JFH-1 confers J6CF replication competence. In this study, we aimed to identify the minimum modifications within these genomic regions needed to establish replication-competent J6CF. We previously identified 4 mutations in the NS5B-to-3=X region that could be used instead of replacement of this region to confer J6CF replication competence. Here, we induced cell cultureadaptive mutations in J6CF by the long-term culture of J6CF/JFH-1 chimeras composed of JFH-1 NS5B-to-3=X or individual parts of this but not the NS3 helicase region. After 2 months of culture, efficient HCV replication and infectious virus production in chimeric RNA-transfected cells were observed, and several amino acid mutations in NS4A were identified in replicating HCV genomes. The introduction of NS4A mutations into the J6CF/JFH-1 chimeras enhanced viral replication and infectious virus production. Immunofluorescence microscopy demonstrated that some of these mutations altered the subcellular localization of the coexpressed NS3 protein and affected the interaction between NS3 and NS4A. Finally, introduction of the most effective NS4A mutation, A1680E, into J6CF contributed to its replication competence in cultured cells when introduced in conjunction with four previously identified adaptive mutations in the NS5B-to-3=X region. In conclusion, we identified an adaptive mutation in NS4A that confers J6CF replication competence when introduced in conjunction with 4 mutations in NS5B-to-3=X and established a replicationcompetent J6CF strain with minimum essential modifications in cultured cells. IMPORTANCEThe HCV cell culture system using the JFH-1 strain and HuH-7 cells can be used to assess the complete HCV life cycle in cultured cells. This cell culture system has been used to develop direct-acting antivirals against HCV, and the ability to use various HCV strains within this system is important for future studies. In this study, we aimed to establish a novel HCV cell culture system using another HCV genotype 2a strain, J6CF, which replicates in chimpanzees but not in cultured cells. We identified an effective cell culture-adaptive mutation in NS4A and established a replication-competent J6CF strain in cultured cells with minimum essential modifications. The described strategy can be used in establishing a novel HCV cell culture system, and the replication-competent J6CF clone composed of the minimum essential modifications needed for cell culture adaptation will be valuable as another representative of genotype 2a strains.

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“…Анализируя образцы крови взрослых больных, M. BouvierAlias и соавторы пришли к выводу, что один фмоль/мл core-антигена соответствует диапазону значений РНК 300-720 МЕ/мл [21], а R.S. Ross и соавторы -конкрет-ному значению 257 МЕ/мл РНК [22]. Ни для детей, ни для взрослых больных нашей страны медианы соотноше-ний РНК ВГС и core-антигена не установлены.…”

Section: результаты и их обсуждениеunclassified

“…Ранее было показано влияние субтипов ВГС на соот-ношение вирусной РНК и core-антигена [22,23]. В на-стоящем исследовании это было изучено для доминирую-щих субтипов вируса у детей с ХГС (табл.…”

Section: результаты и их обсуждениеunclassified

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Investigation of the Significance of Detection of the Nucleocapsid Antigen of the Hepatitis C Virus in Children With Chronic Hepatitis C

Лейбман¹,

Николаева²,

Сапронов³

et al. 2017

Det. infekc.

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Japanese Reference Panel of Blood Specimens for Evaluation of Hepatitis C Virus RNA and Core Antigen Quantitative Assays

Cited by 38 publications

References 34 publications

Hepatitis C Virus Core Antigen: A Simplified Treatment Monitoring Tool, including for Post-Treatment Relapse

Hepatitis C Virus Core Antigen: A Simplified Treatment Monitoring Tool, including for Post-Treatment Relapse

Amino Acid Mutations in the NS4A Region of Hepatitis C Virus Contribute to Viral Replication and Infectious Virus Production

Investigation of the Significance of Detection of the Nucleocapsid Antigen of the Hepatitis C Virus in Children With Chronic Hepatitis C

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