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1.
Gut ; 62(9): 1340-6, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-23135762

RESUMEN

OBJECTIVE: Recent studies have demonstrated that genetic polymorphisms near the IL28B gene are associated with the clinical outcome of pegylated interferon α (peg-IFN-α) plus ribavirin therapy for patients with chronic hepatitis C virus (HCV). However, it is unclear whether genetic variations near the IL28B gene influence hepatic interferon (IFN)-stimulated gene (ISG) induction or cellular immune responses, lead to the viral reduction during IFN treatment. DESIGN: Changes in HCV-RNA levels before therapy, at day 1 and weeks 1, 2, 4, 8 and 12 after administering peg-IFN-α plus ribavirin were measured in 54 patients infected with HCV genotype 1. Furthermore, we prepared four lines of chimeric mice having four different lots of human hepatocytes containing various single nucleotide polymorphisms (SNP) around the IL28B gene. HCV infecting chimeric mice were subcutaneously administered with peg-IFN-α for 2 weeks. RESULTS: There were significant differences in the reduction of HCV-RNA levels after peg-IFN-α plus ribavirin therapy based on the IL28B SNP rs8099917 between TT (favourable) and TG/GG (unfavourable) genotypes in patients; the first-phase viral decline slope per day and second-phase slope per week in TT genotype were significantly higher than in TG/GG genotype. On peg-IFN-α administration to chimeric mice, however, no significant difference in the median reduction of HCV-RNA levels and the induction of antiviral ISG was observed between favourable and unfavourable human hepatocyte genotypes. CONCLUSIONS: As chimeric mice have the characteristic of immunodeficiency, the response to peg-IFN-α associated with the variation in IL28B alleles in chronic HCV patients would be composed of the intact immune system.


Asunto(s)
Hepacivirus/efectos de los fármacos , Hepatitis C Crónica , Interferón-alfa/farmacología , Interleucinas/genética , Polietilenglicoles/farmacología , ARN Viral/análisis , Ribavirina/farmacología , Animales , Antivirales/farmacología , Quimioterapia Combinada , Femenino , Hepatitis C Crónica/tratamiento farmacológico , Hepatitis C Crónica/genética , Humanos , Inmunidad Celular/efectos de los fármacos , Inmunidad Celular/genética , Interferones , Masculino , Ratones , Ratones SCID , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , Proteínas Recombinantes/farmacología , Factores de Tiempo , Resultado del Tratamiento , Carga Viral/métodos
2.
Hum Mol Genet ; 20(17): 3507-16, 2011 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-21659334

RESUMEN

Hematologic abnormalities during current therapy with pegylated interferon and ribavirin (PEG-IFN/RBV) for chronic hepatitis C (CHC) often necessitate dose reduction and premature withdrawal from therapy. The aim of this study was to identify host factors associated with IFN-induced thrombocytopenia by genome-wide association study (GWAS). In the GWAS stage using 900K single-nucleotide polymorphism (SNP) microarrays, 303 Japanese CHC patients treated with PEG-IFN/RBV therapy were genotyped. One SNP (rs11697186) located on DDRGK1 gene on chromosome 20 showed strong associations in the minor-allele-dominant model with the decrease of platelet counts in response to PEG-IFN/RBV therapy [P = 8.17 × 10(-9); odds ratio (OR) = 4.6]. These associations were replicated in another sample set (n = 391) and the combined P-values reached 5.29 × 10(-17) (OR = 4.5). Fine mapping with 22 SNPs around DDRGK1 and ITPA genes showed that rs11697186 at the GWAS stage had a strong linkage disequilibrium with rs1127354, known as a functional variant in the ITPA gene. The ITPA-AA/CA genotype was independently associated with a higher degree of reduction in platelet counts at week 4 (P < 0.0001), as well as protection against the reduction in hemoglobin, whereas the CC genotype had significantly less reduction in the mean platelet counts compared with the AA/CA genotype (P < 0.0001 for weeks 2, 4, 8, 12), due to a reactive increase of the platelet count through weeks 1-4. Our present results may provide a valuable pharmacogenetic diagnostic tool for tailoring PEG-IFN/RBV dosing to minimize drug-induced adverse events.


Asunto(s)
Antivirales/uso terapéutico , Estudio de Asociación del Genoma Completo/métodos , Hepatitis C Crónica/tratamiento farmacológico , Interferones/uso terapéutico , Pirofosfatasas/genética , Ribavirina/uso terapéutico , Trombocitopenia/genética , Antivirales/efectos adversos , Genotipo , Humanos , Interferones/efectos adversos , Desequilibrio de Ligamiento/genética , Polimorfismo de Nucleótido Simple/genética , Ribavirina/efectos adversos , Trombocitopenia/inducido químicamente
3.
J Clin Microbiol ; 51(11): 3484-91, 2013 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-23946517

RESUMEN

We modified and automated a highly sensitive chemiluminescent enzyme immunoassay (CLEIA) for surface antigen (HBsAg) detection using a combination of monoclonal antibodies, each for a specific epitope of HBsAg, and by improving an earlier conjugation technique. Of 471 hepatitis B virus (HBV) carriers seen in our hospital between 2009 and 2012, 26 were HBsAg seronegative as determined by the Abbott Architect assay. The Lumipulse HBsAg-HQ assay was used to recheck those 26 patients who demonstrated seroclearance by the Abbott Architect assay. The performance of the Lumipulse HBsAg-HQ assay was compared with that of a quantitative HBsAg detection system (Abbott Architect) and the Roche Cobas TaqMan HBV DNA assay (CTM) (lower limit of detection, 2.1 log copies/ml) using blood serum samples from patients who were determined to be HBsAg seronegative by the Abbott Architect assay. Ten patients had spontaneous HBsAg loss. Of 8 patients treated with nucleotide analogues (NAs), two were HBsAg seronegative after stopping lamivudine therapy and 6 were HBsAg seronegative during entecavir therapy. Eight acute hepatitis B (AH) patients became HBsAg seronegative. Of the 26 patients, 16 were HBsAg positive by the Lumipulse HBsAg-HQ assay but negative by the Abbott Architect assay. The differences between the two assays in terms of detectable HBsAg persisted over the long term in the spontaneous loss group (median, 10 months), the NA-treated group (2.5 months), and the AH group (0.5 months). In 9 patients, the Lumipulse HBsAg-HQ assay detected HBsAg when HBV DNA was negative by the CTM assay. HBsAg was also detected by the Lumipulse HBsAg-HQ assay in 4 patients with an anti-HBs concentration of >10 mIU/ml, 3 of whom had no HBsAg escape mutations. The automatic, highly sensitive HBsAg CLEIA Lumipulse HBsAg-HQ is a convenient and precise assay for HBV monitoring.


Asunto(s)
Técnicas de Laboratorio Clínico/métodos , Antígenos de Superficie de la Hepatitis B/sangre , Hepatitis B/diagnóstico , Adulto , Anciano , Antivirales/uso terapéutico , Automatización de Laboratorios/métodos , Femenino , Hepatitis B/tratamiento farmacológico , Humanos , Técnicas para Inmunoenzimas/métodos , Mediciones Luminiscentes , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad , Suero/química
4.
J Hepatol ; 56(3): 602-8, 2012 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-22027574

RESUMEN

BACKGROUND & AIMS: Assessment of the risk of hepatocellular carcinoma (HCC) development is essential for formulating personalized surveillance or antiviral treatment plan for chronic hepatitis C. We aimed to build a simple model for the identification of patients at high risk of developing HCC. METHODS: Chronic hepatitis C patients followed for at least 5 years (n=1003) were analyzed by data mining to build a predictive model for HCC development. The model was externally validated using a cohort of 1072 patients (472 with sustained virological response (SVR) and 600 with nonSVR to PEG-interferon plus ribavirin therapy). RESULTS: On the basis of factors such as age, platelet, albumin, and aspartate aminotransferase, the HCC risk prediction model identified subgroups with high-, intermediate-, and low-risk of HCC with a 5-year HCC development rate of 20.9%, 6.3-7.3%, and 0-1.5%, respectively. The reproducibility of the model was confirmed through external validation (r(2)=0.981). The 10-year HCC development rate was also significantly higher in the high-and intermediate-risk group than in the low-risk group (24.5% vs. 4.8%; p<0.0001). In the high-and intermediate-risk group, the incidence of HCC development was significantly reduced in patients with SVR compared to those with nonSVR (5-year rate, 9.5% vs. 4.5%; p=0.040). CONCLUSIONS: The HCC risk prediction model uses simple and readily available factors and identifies patients at a high risk of HCC development. The model allows physicians to identify patients requiring HCC surveillance and those who benefit from IFN therapy to prevent HCC.


Asunto(s)
Carcinoma Hepatocelular/epidemiología , Minería de Datos/métodos , Hepatitis C Crónica/epidemiología , Neoplasias Hepáticas/epidemiología , Modelos Estadísticos , Adulto , Anciano , Antivirales/uso terapéutico , Estudios de Cohortes , Minería de Datos/estadística & datos numéricos , Árboles de Decisión , Femenino , Hepatitis C Crónica/tratamiento farmacológico , Humanos , Interferones/uso terapéutico , Masculino , Persona de Mediana Edad , Polietilenglicoles/uso terapéutico , Ribavirina/uso terapéutico , Medición de Riesgo/métodos , Factores de Riesgo
5.
Clin Infect Dis ; 52(5): 624-32, 2011 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-21245155

RESUMEN

BACKGROUND: Studies of the transmissibility of hepatitis B virus (HBV) in occult hepatitis B (OHB) through blood transfusion are scarce. We aimed to determine the transmissibility of HBV in blood donors with OHB through transfusion in animal and human studies. METHODS: Among 217,595 blood donors, 67 donors with OHB were identified. Four chimeric mice populated with human hepatocytes were inoculated with 2 donor serum samples. Serial serum and liver HBV DNA levels were measured. Forty-nine recipients of blood transfusions traced from 10 donors with OHB (9 of whom were positive for antibody to hepatitis B surface antigen [anti-HBs]) were tested for HBV infection. Homology and phylogenetic analyses between the HBV genomic sequences of donors and recipients were performed. RESULTS: Serum HBV DNA was detectable (10(4) copies/mL) in 1 mouse at weeks 5 and 7 after inoculation. Total HBV DNA and HBV replication template (covalently closed circular DNA) and hepatitis B core antigen were detected in the mouse liver. After transfusion, 45 recipients (91.8%) had no HBV infection (ie, they tested negative for hepatitis B surface antigen and HBV DNA). Four tested positive for HBV DNA. In 3 recipients, 83%-86% homology and distant phylogenetic relatedness with their donor HBV excluded transmission through transfusion. The remaining recipient HBV had 95% sequence homology with her donor HBV, compatible with acquisition of HBV infection from the transfusion. High anti-HBs levels in 7 other recipients suggested recent transfusion-related HBV immune response. CONCLUSIONS: OHB donor blood infectivity was shown in our animal and human studies. However, the risk of HBV transmission in humans was low, especially from blood products obtained from donors with OHB who were anti-HBs positive.


Asunto(s)
Sangre/virología , Virus de la Hepatitis B/aislamiento & purificación , Hepatitis B/transmisión , Reacción a la Transfusión , Adolescente , Adulto , Animales , Donantes de Sangre , ADN Viral/genética , ADN Viral/aislamiento & purificación , Modelos Animales de Enfermedad , Femenino , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/genética , Humanos , Hígado/virología , Masculino , Ratones , Persona de Mediana Edad , Filogenia , Medición de Riesgo , Análisis de Secuencia de ADN , Adulto Joven
6.
J Hepatol ; 54(3): 439-48, 2011 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-21129805

RESUMEN

BACKGROUND & AIMS: Pegylated interferon and ribavirin (PEG-IFN/RBV) therapy for chronic hepatitis C virus (HCV) genotype 1 infection is effective in 50% of patients. Recent studies revealed an association between the IL28B genotype and treatment response. We aimed to develop a model for the pre-treatment prediction of response using host and viral factors. METHODS: Data were collected from 496 patients with HCV genotype 1 treated with PEG-IFN/RBV at five hospitals and universities in Japan. IL28B genotype and mutations in the core and IFN sensitivity determining region (ISDR) of HCV were analyzed to predict response to therapy. The decision model was generated by data mining analysis. RESULTS: The IL28B polymorphism correlated with early virological response and predicted null virological response (NVR) (odds ratio=20.83, p<0.0001) and sustained virological response (SVR) (odds ratio=7.41, p<0.0001) independent of other covariates. Mutations in the ISDR predicted relapse and SVR independent of IL28B. The decision model revealed that patients with the minor IL28B allele and low platelet counts had the highest NVR (84%) and lowest SVR (7%), whereas those with the major IL28B allele and mutations in the ISDR or high platelet counts had the lowest NVR (0-17%) and highest SVR (61-90%). The model had high reproducibility and predicted SVR with 78% specificity and 70% sensitivity. CONCLUSIONS: The IL28B polymorphism and mutations in the ISDR of HCV were significant pre-treatment predictors of response to PEG-IFN/RBV. The decision model, including these host and viral factors may support selection of optimum treatment strategy for individual patients.


Asunto(s)
Hepacivirus/genética , Hepatitis C Crónica/tratamiento farmacológico , Hepatitis C Crónica/genética , Interferón Tipo I/uso terapéutico , Interleucinas/genética , Polimorfismo de Nucleótido Simple , Ribavirina/uso terapéutico , Anciano , Alelos , Antivirales/uso terapéutico , Minería de Datos , Árboles de Decisión , Farmacorresistencia Viral/genética , Femenino , Genes Virales , Genotipo , Hepacivirus/efectos de los fármacos , Hepatitis C Crónica/inmunología , Hepatitis C Crónica/virología , Humanos , Interferones , Japón , Modelos Logísticos , Masculino , Persona de Mediana Edad , Modelos Biológicos , Mutación , Pronóstico , ARN Viral/sangre , ARN Viral/genética , Proteínas Recombinantes , Estudios Retrospectivos , Resultado del Tratamiento , Proteínas del Núcleo Viral/genética , Carga Viral
7.
J Med Virol ; 83(3): 412-8, 2011 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-21264861

RESUMEN

The virological characteristics of hepatitis B virus (HBV) implicated in the reactivation of occult hepatitis B in patients who have received hematopoietic stem-cell transplantation or chemotherapy for the hematological malignancy are not well defined. Twenty-eight HBsAg-negative patients who received hematopoietic stem-cell transplantation and 138 HBsAg-negative patients treated for malignant lymphoma with chemotherapy including rituximab were enrolled. Three of the 28 patients (10.7%) received hematopoietic stem-cell transplantation and one of the 138 (0.72%) patients treated for malignant lymphoma with chemotherapy developed de novo HBV hepatitis. Anti-HBc was detected in four and anti-HBs in two patients. Genotype Bj was detected in two and C in two of they all possessed wild-type sequences in the core promoter region. A precore stop mutation (A1896) was detected in a patient with genotype Bj who developed fulminant hepatic failure. HBV DNA was detected in pretreatment HBsAg-negative samples in two of four patients, and the HBV genome sequence identified from sera before chemotherapy and at the time of de novo HBV hepatitis showed 100% homology. In an in vitro replication model, genotype Bj with the A1896 clone obtained from a fulminant case had a replication level much higher than clones obtained from de novo hepatitis B patients with genotype Bj or C with G1896. In conclusion, this is the first report demonstrating de novo hepatitis B from the reactivation of occult HBV infection confirmed by molecular evolutional analysis. The fulminant outcome of HBV reactivation can be associated with genotype Bj exhibiting high replication due to the A1896 mutation.


Asunto(s)
Neoplasias Hematológicas/complicaciones , Neoplasias Hematológicas/virología , Virus de la Hepatitis B/fisiología , Hepatitis B/complicaciones , Hepatitis B/virología , Activación Viral , Adulto , Antineoplásicos/uso terapéutico , Línea Celular Tumoral , ADN Viral/análisis , Femenino , Genotipo , Neoplasias Hematológicas/tratamiento farmacológico , Neoplasias Hematológicas/inmunología , Trasplante de Células Madre Hematopoyéticas , Hepatitis B/inmunología , Humanos , Masculino , Persona de Mediana Edad , Mutación , Filogenia , Regiones Promotoras Genéticas , Estudios Retrospectivos
8.
J Med Virol ; 83(3): 445-52, 2011 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-21264865

RESUMEN

The aim of the present study was to clarify the significance of viral factors for pretreatment prediction of sustained virological response to pegylated-interferon (PEG-IFN) plus ribavirin (RBV) therapy for chronic hepatitis C using data mining analysis. Substitutions in the IFN sensitivity-determining region (ISDR) and at position 70 of the HCV core region (Core70) were determined in 505 patients with genotype 1b chronic hepatitis C treated with PEG-IFN plus RBV. Data mining analysis was used to build a predictive model of sustained virological response in patients selected randomly (n = 304). The reproducibility of the model was validated in the remaining 201 patients. Substitutions in ISDR (odds ratio = 9.92, P < 0.0001) and Core70 (odds ratio = 1.92, P = 0.01) predicted sustained virological response independent of other covariates. The decision-tree model revealed that the rate of sustained virological response was highest (83%) in patients with two or more substitutions in ISDR. The overall rate of sustained virological response was 44% in patients with a low number of substitutions in ISDR (0-1) but was 83% in selected subgroups of younger patients (<60 years), wild-type sequence at Core70, and higher level of low-density lipoprotein cholesterol (LDL-C) (≥ 120 mg/dl). Reproducibility of the model was validated (r(2) = 0.94, P < 0.001). In conclusion, substitutions in ISDR and Core70 of HCV are significant predictors of response to PEG-IFN plus RBV therapy. A decision-tree model that includes these viral factors as predictors could identify patients with a high probability of sustained virological response.


Asunto(s)
Antivirales/uso terapéutico , Hepacivirus/genética , Hepatitis C Crónica , Interferón-alfa/uso terapéutico , Polietilenglicoles/uso terapéutico , Ribavirina/uso terapéutico , Proteínas del Núcleo Viral/genética , Estudios de Cohortes , Minería de Datos , Árboles de Decisión , Quimioterapia Combinada , Genotipo , Hepatitis C Crónica/tratamiento farmacológico , Hepatitis C Crónica/genética , Humanos , Interferón alfa-2 , Análisis Multivariante , Mutación , Proteínas Recombinantes/uso terapéutico , Estudios Retrospectivos , Resultado del Tratamiento
9.
Rinsho Byori ; 59(9): 838-43, 2011 Sep.
Artículo en Japonés | MEDLINE | ID: mdl-22111300

RESUMEN

There are 1.5 million hepatitis B virus (HBV) infected patients in Japan. Anti-viral therapy is important for chronic hepatitis B patients to prevent hepatocellular carcinoma. Recently, HBs antigen (HBsAg) quantification has been reported to be useful for not only HBV screening but also for monitoring of anti-viral treatment. In this paper, we evaluated the clinical utility of quantitative assay of HBsAg by HISCL HBsAg kit. Although there can be a significant difference in age, HBeAg positive/negative and viral genotype, there is not in the disease stage. Moreover, the weak correlation was confirmed between HBsAg and HBV-DNA levels with or without anti-virus treatment. In the clinical practice, as HBV-DNA becomes undetectable immediately by anti-viral therapy such as entecavir, it may be difficult to evaluate the efficacy. The monitoring of the HBsAg concentration in addition to HBV-DNA would be useful for the evaluation. Hence, the clinical role of HBsAg concentration could spread widely in Japan.


Asunto(s)
Antígenos de Superficie de la Hepatitis B/sangre , Hepatitis B Crónica/diagnóstico , Adulto , Anciano , Antivirales/uso terapéutico , Biomarcadores/sangre , Carcinoma Hepatocelular/prevención & control , Carcinoma Hepatocelular/virología , ADN Viral/sangre , Genotipo , Virus de la Hepatitis B/genética , Hepatitis B Crónica/tratamiento farmacológico , Humanos , Neoplasias Hepáticas/prevención & control , Neoplasias Hepáticas/virología , Persona de Mediana Edad , Monitoreo Fisiológico , Juego de Reactivos para Diagnóstico
10.
J Infect Dis ; 201(11): 1663-71, 2010 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-20420509

RESUMEN

BACKGROUND: Approximately 20% of patients with hepatitis C virus (HCV) genotype 1b infection have nonresponse to the most current treatment, pegylated interferon with ribavirin. Mutations in the HCV core region were recently proposed to be associated with nonresponse. Our aim was to evaluate the viral factors associated with treatment failure. METHODS: HCV variants were determined directly and after cloning in 66 HCV-1b-infected Japanese patients and in 5 urokinase-type plasminogen activator transgenic severe combined immunodeficiency mice with human hepatocytes (chimeric mice), at baseline, during treatment, and after treatment. RESULTS: At baseline, glutamine at position 70 of the HCV core protein (70Q) was detected by direct sequencing in 20% of patients with virologic response and in 43.8% of patients with nonresponse. Among patients with nonresponse, who were examined during and after treatment, the prevalence of the 70Q substitution increased to 56.3%, which indicates that treatment-induced selection occurred in all patients with nonresponse who had 70Q quasispecies detectable by cloning. This observation was reinforced by the results from experimentally infected chimeric mice. Logistic regression analysis indicated that detection of 70Q quasispecies was associated with a statistically significantly increased risk of nonresponse (odds ratio, 15.1; P = .004) in the studied cohort. CONCLUSION: Presence of the 70Q quasispecies at baseline was associated with an increased risk of treatment failure, as indicated by the positive selection of the 70Q clones induced by treatment with pegylated interferon with ribavirin. These results urge further investigation of the mechanisms of this association.


Asunto(s)
Sustitución de Aminoácidos/genética , Hepacivirus/clasificación , Hepatitis C Crónica/virología , Interferón-alfa/uso terapéutico , Polietilenglicoles/uso terapéutico , Ribavirina/uso terapéutico , Selección Genética , Proteínas del Núcleo Viral/genética , Adulto , Anciano , Animales , Antivirales/uso terapéutico , Femenino , Genotipo , Hepacivirus/genética , Hepacivirus/aislamiento & purificación , Hepatitis C Crónica/tratamiento farmacológico , Humanos , Interferón alfa-2 , Japón , Masculino , Ratones , Ratones Transgénicos , Persona de Mediana Edad , ARN Viral/genética , Proteínas Recombinantes , Análisis de Secuencia de ADN , Insuficiencia del Tratamiento
11.
J Virol ; 83(20): 10538-47, 2009 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-19640977

RESUMEN

Hepatitis B virus (HBV) of a novel genotype (J) was recovered from an 88-year-old Japanese patient with hepatocellular carcinoma who had a history of residing in Borneo during the World War II. It was divergent from eight human (A to H) and four ape (chimpanzee, gorilla, gibbon, and orangutan) HBV genotypes, as well as from a recently proposed ninth human genotype I, by 9.9 to 16.5% of the entire genomic sequence and did not have evidence of recombination with any of the nine human genotypes and four nonhuman genotypes. Based on a comparison of the entire nucleotide sequence against 1,440 HBV isolates reported, HBV/J was nearest to the gibbon and orangutan genotypes (mean divergences of 10.9 and 10.7%, respectively). Based on a comparison of four open reading frames, HBV/J was closer to gibbon/orangutan genotypes than to human genotypes in the P and large S genes and closest to Australian aboriginal strains (HBV/C4) and orangutan-derived strains in the S gene, whereas it was closer to human than ape genotypes in the C gene. HBV/J shared a deletion of 33 nucleotides at the start of preS1 region with C4 and gibbon genotypes, had an S-gene sequence similar to that of C4, and expressed the ayw subtype. Efficient infection, replication, and antigen expression by HBV/J were experimentally established in two chimeric mice with the liver repopulated for human hepatocytes. The HBV DNA sequence recovered from infected mice was identical to that in the inoculum. Since HBV/J is positioned phylogenetically in between human and ape genotypes, it may help to trace the origin of HBV and merits further epidemiological surveys.


Asunto(s)
Enfermedades del Simio Antropoideo/virología , Evolución Molecular , Variación Genética , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/genética , Hepatitis B Crónica/virología , Anciano de 80 o más Años , Secuencia de Aminoácidos , Animales , Borneo , ADN Viral/análisis , Genotipo , Virus de la Hepatitis B/aislamiento & purificación , Humanos , Hylobates , Masculino , Ratones , Datos de Secuencia Molecular , Pan troglodytes , Filogenia , Pongo pygmaeus , Análisis de Secuencia de ADN
12.
J Med Virol ; 82(11): 1878-88, 2010 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-20872714

RESUMEN

Accuracy for monitoring of the concentration of hepatitis C virus (HCV) RNA represents a major challenge throughout the management of patients with chronic hepatitis C. To investigate the genotype-independent efficiency and the accuracy of two real-time detection reverse transcription-polymerase chain reaction (RT-PCR) assays; the Cobas Ampliprep/Cobas TaqMan (CAP/CTM); and the Abbott RealTime HCV (ART), a total of 184 samples with different HCV subtypes were examined; 1b (n=58), 2a (n=39), 2b (n=26), 3a (n=20), and 4 (n=41). A robust linear correlation was observed between the two assays applied to genotypes 1b, 2a, 2b, and 3a [the correlation coefficient (R) ranged from 0.99 to 0.98], but not to genotype 4 specimens (R=0.78). A significant difference in measurements of HCV RNA using CAP/CTM and ART in serum samples with genotypes 1b and 4 was observed (0.72, -0.53 log IU/ml, P<0.0001, 0.01, respectively). A robust correlation was observed between the HCV core antigen and HCV RNA values by either of the HCV RNA quantitation assays applied to all genotypes with exception of genotype 4, for which R was higher with ART (R=0.95) than with CAP/CTM (R=0.80). The lower limit of detection of CAP/CTM and ART were 41.4 and 28.5 IU/ml using the WHO standards, respectively. In conclusion, two RT-PCR assays had a high efficiency and accuracy for quantitation of HCV RNA of genotypes 2a, 2b, and 3a, but the mean values of HCV RNA differed for genotype 1b and 4.


Asunto(s)
Hepacivirus/clasificación , Hepacivirus/genética , Hepatitis C Crónica/virología , ARN Viral/sangre , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/normas , Regiones no Traducidas 5'/genética , Genoma Viral , Genotipo , Hepacivirus/aislamiento & purificación , Antígenos de la Hepatitis C/sangre , Hepatitis C Crónica/diagnóstico , Humanos , Inmunoensayo , Luminiscencia , Datos de Secuencia Molecular , ARN Viral/análisis , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Análisis de Secuencia de ADN , Proteínas del Núcleo Viral/sangre
13.
J Clin Microbiol ; 47(2): 385-9, 2009 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-19091819

RESUMEN

Two commercial real-time PCR assays are currently available for sensitive hepatitis C virus (HCV) RNA quantification: the Abbott RealTime HCV assay (ART) and Roche Cobas AmpliPrep/Cobas TaqMan HCV assay (CAP/CTM). We assessed whether the two real-time PCR assays were more effective than Roche Cobas Amplicor HCV Monitor test, v.2.0 (CAM) for prediction of the sustained virological response (SVR) to pegylated interferon (PEG-IFN) plus ribavirin (RBV) in chronic hepatitis C. Sixty patients chronically infected with HCV genotype 1b (37 males and 23 females, 53 +/- 12 years of age) were treated with PEG-IFNalpha2b plus RBV for 48 weeks. Stored specimens at nine time points for each patient (at baseline, on treatment, and 24 weeks after treatment) were tested by the two real-time PCR assays and CAM. Twenty-six (43.3%) patients reached SVR. The positive predictive values (PPVs) for SVR of undetectable HCV RNA at week 12 by CAM, ART, and CAP/CTM were 74.3%, 88.0%, and 95.2%, respectively. An undetectable HCV RNA level by CAM, ART, and CAP/CTM correctly predicted SVR at week 4 in 100%, 100%, and 100% of patients, at weeks 5 to 8 in 91.7%, 100%, and 100% of patients, at weeks 9 to 12 in 55.6%, 75%, and 87.5% of patients, and at weeks 13 to 24 in 0%, 26.7%, and 40% of patients, respectively. Of 16 patients who relapsed after treatment, HCV RNA was detectable in 2 patients at the end of treatment by CAP/CTM but undetectable by ART and CAM. HCV RNA tests using ART and CAP/CTM are considered to be more effective at predicting SVR than CAM, and the PPV for SVR was slightly higher in CAP/CTM than in ART.


Asunto(s)
Antivirales/uso terapéutico , Sangre/virología , Hepacivirus/aislamiento & purificación , Hepatitis C Crónica/tratamiento farmacológico , Interferón-alfa/uso terapéutico , Reacción en Cadena de la Polimerasa/métodos , Ribavirina/uso terapéutico , Adulto , Anciano , Femenino , Hepacivirus/genética , Hepatitis C Crónica/virología , Humanos , Interferón alfa-2 , Masculino , Persona de Mediana Edad , Polietilenglicoles , Proteínas Recombinantes , Factores de Tiempo
14.
J Clin Microbiol ; 47(5): 1476-83, 2009 May.
Artículo en Inglés | MEDLINE | ID: mdl-19297602

RESUMEN

Acute hepatitis B virus (HBV) infection has been increasing through promiscuous sexual contacts, and HBV genotype A (HBV/A) is frequent in patients with acute hepatitis B (AHB) in Japan. To compare the geographic distribution of HBV genotypes in patients with chronic hepatitis B (CHB) in Japan between 2005 and 2006 and between 2000 and 2001, with special attention to changes in the proportion of HBV/A, a cohort study was performed to survey changes in genotypes of CHB patients at 16 hospitals throughout Japan. Furthermore, we investigated the clinical characteristics of each genotype and examined the genomic characteristics of HBV/A isolates by molecular evolutionary analyses. Of the 1,271 patients, 3.5%, 14.1%, and 82.3% were infected with HBV/A, -B, and -C, respectively. In comparison with our previous survey during 2000 and 2001, HBV/A was twice as frequent (3.5% versus 1.7%; P = 0.02). The mean age was lower in the patients with HBV/A than in those with HBV/B or -C. Based on phylogenetic analyses of 11 full-length genomes and 29 pre-S2/S region sequences from patients, HBV/A isolates were imported from Europe and the United States, as well as the Philippines and India. They clustered with HBV/A from AHB patients and have spread throughout Japan. HBV/A has been increasing in CHB patients in Japan as a consequence of AHB spreading in the younger generation through promiscuous sexual contacts, aided by a tendency of HBV/A to induce chronic hepatitis. The spread of HBV/A infection in Japan should be prevented by universal vaccination programs.


Asunto(s)
ADN Viral/genética , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/genética , Hepatitis B Crónica/virología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Análisis por Conglomerados , Femenino , Genotipo , Virus de la Hepatitis B/aislamiento & purificación , Humanos , Japón , Masculino , Persona de Mediana Edad , Epidemiología Molecular , Datos de Secuencia Molecular , Filogenia , Análisis de Secuencia de ADN , Adulto Joven
15.
J Med Virol ; 81(6): 1015-23, 2009 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-19382263

RESUMEN

Egypt is one of the countries with very high rates of hepatitis C virus (HCV) related morbidity and mortality. However, little is known about geographical and clinical differences in genetic variability of HCV in Egypt. Using direct sequencing and phylogenetic analysis of partial core/E1 and NS5B regions of the HCV genome, HCV genotype/subtype was determined in 129 HCV-infected patients residing in three governates in south Egypt: Assuit, Sohag, and Qena. According to clinical stage of infection, patients were categorized into four groups: asymptomatic carriers, n = 16; chronic hepatitis C patients, n = 36; liver cirrhosis, n = 54; and hepatocellular carcinoma (HCC), n = 23. Genotype 4a was detected in 80.6%, whereas 1g, 4l, 4n, 4o, 4f, and 4m were identified in 7.7%, 4.7%, 3.9%, 1.6%, 0.8%, and 0.8% of cases, respectively. The prevalence of 4a differed regionally; from 88.5% (in Sohag) to 64% (in Assuit, P = 0.002). Genotypes 4l and 4n had a higher prevalence in Assuit (12.8%, 10.3%) than Sohag (0%, 0%; P < or = 0.011). Difference in clinical features of determined genotypes/subtypes was observed; more carriers of non-4a variants (4l and 4n, 4f, or 4m) had chronic hepatitis compared to carriers of 4a (53.3% vs. 23.1%, P = 0.025), while more patients with 4a had liver cirrhosis (45.2% vs. 13.3%, P = 0.023). Two HCV-4o strains were isolated in this study, both from patients with HCC. In conclusion, geographical diversity of HCV was revealed in this study in southern Egypt. A further case-control study is required to confirm the trends of differential pathogenicity of HCV subtypes, indicated by this study.


Asunto(s)
Hepacivirus/clasificación , Hepacivirus/genética , Hepatitis C/patología , Hepatitis C/virología , Polimorfismo Genético , Adulto , Anciano , Egipto , Femenino , Genotipo , Hepacivirus/aislamiento & purificación , Hepacivirus/patogenicidad , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Filogenia , Análisis de Secuencia de ADN , Índice de Severidad de la Enfermedad , Proteínas del Núcleo Viral/genética , Proteínas del Envoltorio Viral/genética , Proteínas no Estructurales Virales/genética , Adulto Joven
16.
Hepatol Res ; 39(1): 21-30, 2009 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-18721155

RESUMEN

AIM: Many reports have revealed ursodeoxycholic acid (UDCA) to be effective against chronic hepatitis C virus (HCV). However, some cases resist this therapy and the mechanism of action remains unclear. In this study, UDCA was administered to patients with chronic HCV and the correlation between the bile acids of the biliary bile and serum and the drug efficacy was investigated. METHODS: Fifteen patients were given 600 mg/day of UDCA for more than 24 weeks. The serum bile acid concentrations and biliary and serum bile acid were collected before and after 24 weeks of UDCA treatment, and composition determined by high-performance liquid chromatography. RESULTS: The treatment was effective in nine cases (ALT decreased to less than twice the normal values 80 IU/L) and ineffective in six cases. There was no significant difference in the serum bile acid concentrations before and after UDCA treatment between the values of both cases. After UDCA treatment, the serum percentage of UDCA (effective, 62.5 +/- 2.0; ineffective, 53.5 +/- 2.5, (P = 0.02)) and the percentage of chenodeoxycholic acid (CDCA) showed no remarkable changes. In the biliary bile the percentage of CDCA (effective, 30.9 +/- 2.0; ineffective, 20.0 +/- 3.0, (P = 0.007)) and the percentage of UDCA showed no remarkable changes. CONCLUSION: In the effective cases, the percentage of UDCA in the serum and the percentage of CDCA in biliary bile were significantly higher than in the ineffective cases. This indicates that, when effective, CDCA decreases in hepatocytes and this reduction contributes to hepatoprotection.

17.
Hepatol Res ; 39(7): 648-56, 2009 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-19456899

RESUMEN

BACKGROUND: Host and viral factors can promote the development of fulminant hepatitis B (FHB), but there have been no case-control studies for figuring out virological parameters that can distinguish FHB. METHODS: In a case-control study, virological factors associated with the development of FHB were sought in 50 patients with FH developed by transient hepatitis B virus (HBV) infection (FH-T) and 50 with acute self-limited hepatitis B (AHB) who were matched for sex and age. In addition, 12 patients with FH developed by acute exacerbation (AE) of asymptomatic HBV carrier (ASC) (FH-C) were also compared with 12 patients without FH by AE of chronic hepatitis B (AE-C). RESULTS: Higher HBV DNA levels, subgenotype B1/Bj, A1762T/G1764A, G1896A, G1899A and A2339G mutation were significantly more frequent (P < 0.05), while hepatitis B e-antigen was less frequent in the FH-T patients than AHB. In multivariate analysis, G1896A mutation (odds ratio [OR], 13.53; 95% confidence interval [CI], 2.75-66.64), serum HBV DNA more than 5.23 log copies/mL (OR, 5.14; 95% CI, 1.10-24.15) and total bilirubin more than 10.35 mg/mL (OR, 7.81; 95% CI, 1.77-34.51) were independently associated with a fulminant outcome by transient HBV infection. On the other hand, in comparison with the patients between FH-C and AE-C groups, there was no significant difference of virological factors associated with the development of FHB. CONCLUSION: A number of virological factors have been defined that may distinguish FH-T from AHB in a case-control study. The pathogenic mechanism of FHB between transient HBV infection and AE of ASC would be different.

18.
Rinsho Byori ; 57(1): 42-7, 2009 Jan.
Artículo en Japonés | MEDLINE | ID: mdl-19227189

RESUMEN

Clinical significance of Hepatitis B virus(HBV) genotyping is increasingly recognized. The aim of this study was to evaluate reproducibility, accuracy, and sensitivity of an enzyme immunoassay (EIA) based HBV genotyping kit, which designed to discriminate between genotypes to A, B, C, or D by detecting genotype-specific epitopes in PreS2 region. Using the four genotypes panels, the EIA demonstrated complete inter and intra-assay genotyping reproducibility. Serum specimens had stable results after 8 days at 4 degrees C, or 10 cycles of freezing-thawing. In 91 samples that have been genotyped by DNA sequencing, 87(95.6%) were in complete accordance with EIA genotyping. Of examined 344 HBsAg-positive serum specimens, genotypes A, B, C and D were determined in 26 (7.6%), 62 (18.0%), 228 (66.3%), and 9 (2.6%) cases, respectively. Of 19 (5.5%) specimens unclassified by the EIA, 13 were found to have low titer of HBsAg concentration (< 3 IU/ml), and the other 5 had amino acid mutations or deletions within targeted PreS2 epitopes. The EIA allowed genotyping even in HBV DNA negative samples (96.2%). In conclusion, HBV genotype EIA is reliable, sensitive and easy assay for HBV genotyping. The assay would be useful for clinical use.


Asunto(s)
Virus de la Hepatitis B/genética , Técnicas para Inmunoenzimas/instrumentación , Genotipo , Virus de la Hepatitis B/inmunología , Técnicas para Inmunoenzimas/normas , Reproducibilidad de los Resultados
19.
Rinsho Byori ; 57(3): 200-5, 2009 Mar.
Artículo en Japonés | MEDLINE | ID: mdl-19363989

RESUMEN

Measurements of serum concentrations of Des-gamma-carboxy Prothrombin (PIVKA-II) are widely used for diagnosing hepatocellular carcinoma (HCC). Recently, in Lumipulsef assay, it was reported that antibodies against alkaline phosphatase (ALP) derived from anti bleeding sheets led false high values of PIVKA-II in the patients with HCC resection. To improve the previous issue, newly developed Lumipulse PrestoII assay was examined. (1) The assay was reliable and positively correlated with the previous assays (Lumipulse f and Picolumi, R = 0.997 and 0.994 (n=115), respectively). (2) Eleven cases, which had false high values of PIVKA-II by the Lumipulsef assay, were examined by the PrestoII assay with excess of inactive ALP. The false high values of 10 cases were improved, but only one was still high. False reactivity of this case was stronger than other cases, more effective adsorption was required. (3) Comparing the absorbent activity of inactive ALP among 6 different kinds, we found inactive ALP with much higher adsorbent activity. When this inactive ALP was applied to assay, false high values of PIVKA-II were improved in all 11 cases. In conclusion, the PrestoII assay, which applies the inactive ALP with high activity, is reliable and useful for clinical screening.


Asunto(s)
Biomarcadores de Tumor/sangre , Biomarcadores/sangre , Carcinoma Hepatocelular/diagnóstico , Neoplasias Hepáticas/diagnóstico , Mediciones Luminiscentes/métodos , Precursores de Proteínas/sangre , Reacciones Falso Positivas , Humanos , Protrombina , Reproducibilidad de los Resultados
20.
Hepatol Res ; 38(5): 457-64, 2008 May.
Artículo en Inglés | MEDLINE | ID: mdl-18034826

RESUMEN

AIM: A series of recent studies have indicated the presence of natural intergenotypic recombinant hepatitis C virus (HCV) strains in distinct parts of the world. The majority of the current genotyping methods are based on analysis of either 5'UTR, structural (Core/E1/E2) or non-structural (NS5B) genomic regions of the virus. METHODS: In the present study, based on both structural and non-structural regions, we determined the genotype of 55 anti-HCV-positive intravenous drug users (IDUs) in Uzbekistan. RESULTS: HCV-3a (67.3%) was the most prevalent genotype in this cohort, followed by HCV-1b (27.3%). A discrepancy in results was observed between structural and non-structural regions in one case (1.8%). Phylogenetically this strain was related to the previously reported RF1_2k/1b variant. Based on accumulated sequences, specific primers were designed for polymerase chain reaction (PCR) spanning the tentative intergenotypic crossover point of RF1_2k/1b. The sensitivity and specificity of the method were assessed using generated template clones of HCV-1b, 2a, 2 k and RF1_2k/1b. The method was applied to 55 cases in the present study and only one case showed a positive result, indicating that in these individuals, the variant is not present as a minor quasispecies clone. CONCLUSION: In conclusion, the finding of RF1_2k/1b in Central Asia indicates that the variant has wide geographic distribution. The PCR-based screening method developed in this study should be useful in further epidemiological and clinical studies on the recombination phenomenon in HCV.

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