Typing of hepatitis C virus isolates and characterization of new subtypes using a line probe assay

Stuyver, Lieven; Rossau, R.; Wyseur, Ann; Duhamel, M; Vanderborght, Bart; Heuverswyn, Hugo Van; Maertens, Geert

doi:10.1099/0022-1317-74-6-1093

Cited by 685 publications

(424 citation statements)

References 25 publications

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“…In this state, we presently found genotypes 1, 2 and 3 at frequencies similar to those previously reported (19)(20)(21)(22), except for genotype 2. In another study involving intravenous drug users from Rio de Janeiro, genotype 2 was found at a higher frequency (23).…”

Section: Discussionsupporting

confidence: 75%

Geographic distribution of hepatitis C virus genotypes in Brazil

Campiotto

Pinho

Carrilho

et al. 2005

Braz J Med Biol Res

174

102

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Brazil is a country of continental dimension with a population of different ethnic backgrounds. Thus, a wide variation in the frequencies of hepatitis C virus (HCV) genotypes is expected to occur. To address this point, 1,688 sequential samples from chronic HCV patients were analyzed. HCV-RNA was amplified by the RT-PCR from blood samples collected from 1995 to 2000 at different laboratories located in different cities from all Brazilian States. Samples were collected in tubes containing a gel separator, centrifuged in the site of collection and sent by express mail in a refrigerated container to Laboratório Bioquímico Jardim Paulista, São Paulo, SP, Brazil. HCV-RNA was extracted from serum and submitted to RT and nested PCR using standard procedures. Nested PCR products were submitted to cycle sequencing reactions without prior purification. Sequences were analyzed for genotype determination and the following frequencies were found: 64.9% (1,095) for genotype 1, 4.6% (78) for genotype 2, 30.2% (510) for genotype 3, 0.2% (3) for genotype 4, and 0.1% (2) for genotype 5. The frequencies of HCV genotypes were statistically different among Brazilian regions (P = 0.00017). In all regions, genotype 1 was the most frequent (51.7 to 74.1%), reaching the highest value in the North; genotype 2 was more prevalent in the Center-West region (11.4%), especially in Mato Grosso State (25.8%), while genotype 3 was more common in the South (43.2%). Genotypes 4 and 5 were rarely found and only in the Southeast, in São Paulo State. The present data indicate the need for careful epidemiological surveys throughout Brazil since knowing the frequency and distribution of the genotypes would provide key information for understanding the spread of HCV.

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Section: Discussionsupporting

confidence: 75%

Geographic distribution of hepatitis C virus genotypes in Brazil

Campiotto

Pinho

Carrilho

et al. 2005

Braz J Med Biol Res

174

102

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“…HCV was genotyped by a nested RT-PCR RESULTS assay using universal biotinylated primers in the 5 noncoding region, in which sequence degeneracy was included to allow annealing Twenty-four patients (9%), 11 in the IFN-a2a group and to all HCV genotypes. 20 The RT-PCR products were then hybridized 13 in the IFN-aN1 group, dropped out of the study, 9 because IFNa-N1) showed no biochemical response (Table 2). Seventy-one patients (36 treated with IFNa-2a and 35 treated with IFNa-N1) showed a transient biochemical response.…”

Section: Methodsmentioning

confidence: 99%

“…To avoid potential bias caused by insufficient dose or duranases and undetectable HCV RNA by RT-PCR after 12 tion of therapy, we elected to treat patients for 12 months months of therapy, but only 19 in the first group and 20 and modify the dose according to the alanine transaminase in the second group (total, 39 [17%]) had biochemical (ALT) response. The initial dose was 6 MU.…”

Section: Ifn-an1 Group (Total 59 [25%] Had Normal Transami-mentioning

confidence: 99%

A Prospective, Randomized Trial Comparing Lymphoblastoid to Recombinant Interferon Alfa 2A As Therapy for Chronic Hepatitis C

et al. 1996

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the development of IFN-neutralizing antibodies. [12][13][14] TheoretTo compare the long-term effectiveness and tolerabilically, these risks might be enhanced by long-term adminity of lymphoblastoid interferon (IFN-aN1) and recombiistration of a single type of IFN and mitigated by the adminnant interferon alfa 2a (IFN-a2a) in patients with istration of different types of IFN. Lymphoblastoid IFN chronic hepatitis caused by hepatitis C virus (HCV), 234 (IFN-aN1), a mixture of 22 interferons, 15 was shown to induce consecutive patients with HCV-related chronic hepatitis a response in 40% of nonresponders to recombinant IFN 16,17 were randomized prospectively to receive titrated doses and resulted in a complete response in all patients with hepa-(starting dose Å 6 million units [MU]) of IFN-a2a (n Å titis C who relapsed during treatment with recombinant 118) or IFN-aN1 (n Å 116) for 12 months. HCV RNA was IFN. 14 To assess whether treatment of hepatitis C can be detected by reverse-transcription polymerase chain reimproved by using lymphoblastoid IFN, we have conducted action (RT-PCR), quantified by branched-DNA (bDNA) a prospective randomized comparative trial in which patients assay, and genotyped by reverse hybridization assay.with chronic hepatitis caused by HCV were randomized to Thirty-one patients in the IFN-a2a group and 28 in the either recombinant IFN-a2a or lymphoblastoid IFN-aN1. IFN-aN1 group (total, 59 [25%] had normal transami-To avoid potential bias caused by insufficient dose or duranases and undetectable HCV RNA by RT-PCR after 12 tion of therapy, we elected to treat patients for 12 months months of therapy, but only 19 in the first group and 20and modify the dose according to the alanine transaminase in the second group (total, 39 [17%]) had biochemical (ALT) response. The initial dose was 6 MU. To gain insight and virological responses 12 months after treatment was on the cost-effectiveness of treatment regimens, we analyzed discontinued. The two treatment groups differed in the tolerability and cost of the two treatments, as well as terms of prevalence of major drug-related adverse reacclinical and virological predictors of treatment outcome that tions (23% vs. 37%, P Å .025). The mean total dose per could be used for refining therapy duration or dosage. size made it possible to perform an interim analysis when 50% of the patients had been enrolled. It was planned that the trial would Less than 25% of the patients with chronic hepatitis C be discontinued at that time if the difference in response rates beachieve sustained biochemical responses with undetectable se-tween the two treatments was outside the confidence interval (CI) related to a type I error equal to 0.025 (3.8%-36.2%). and longer duration of therapy have been attempted to in-ies, and abnormal serum transaminase levels. Enrollment was crease the response rates, but the data are inconclusive. [4][5][6][7][8][9] stopped in November 1992 when 258 patients had been recruited, Nonresponse or loss of response during IFN therapy could and...

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“…The procedure was repeated three times. After centrifugation at 10,000 ϫ g for 1 min, the supernatant was subjected to a standard extraction procedure and amplification by a two-round nested RT-PCR as reported previously (27). Firstand second-round PCR products were subjected to electrophoresis on a 2% agarose gel containing ethidium bromide.…”

Section: Binding Of Hvr1-specific Mabs To Bona Fide Hcv Particlesmentioning

confidence: 99%

Monoclonal Antibodies with Broad Specificity for Hepatitis C Virus Hypervariable Region 1 Variants Can Recognize Viral Particles

Cerino

Meola

Segagni

et al. 2001

The Journal of Immunology

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The hypervariable region 1 (HVR1) of the E2 protein of hepatitis C virus (HCV) is a highly heterogeneous sequence that is promiscuously recognized by human sera via binding to amino acid residues with conserved physicochemical properties. We generated a panel of mAbs from mice immunized with HVR1 surrogate peptides (mimotopes) affinity-selected with sera from HCV-infected patients from a phage display library. A high number of specific clones was obtained after immunization with a pool of nine mimotopes, and the resulting mAbs were shown to recognize several 16- and 27-mer peptides derived from natural HVR1 sequences isolated from patients with acute and chronic HCV infection, suggesting that HVR1 mimotopes were efficient antigenic and immunogenic mimics of naturally occurring HCV variants. Moreover, most mAbs were shown to bind HVR1 in the context of a complete soluble form of the E2 glycoprotein, indicating recognition of correctly folded HVR1. In addition, a highly promiscuous mAb was able to specifically capture bona fide viral particles (circulating HCV RNA) as well as rHCV-like particles assembled in insect cells expressing structural viral polypeptides derived from an HCV 1a isolate. These findings demonstrate that it is possible to induce a broadly cross-reactive clonal Ab response to multiple HCV variants. In consideration of the potentially important role of HVR1 in virus binding to cellular receptor(s), such a mechanism could be exploited for induction of neutralizing Abs specific for a large repertoire of viral variants.

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Typing of hepatitis C virus isolates and characterization of new subtypes using a line probe assay

Cited by 685 publications

References 25 publications

Geographic distribution of hepatitis C virus genotypes in Brazil

Geographic distribution of hepatitis C virus genotypes in Brazil

A Prospective, Randomized Trial Comparing Lymphoblastoid to Recombinant Interferon Alfa 2A As Therapy for Chronic Hepatitis C

Monoclonal Antibodies with Broad Specificity for Hepatitis C Virus Hypervariable Region 1 Variants Can Recognize Viral Particles

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