Sequencing of E2 and NS5A regions of HCV genotype 3a in Brazilian patients with chronic hepatitis

Hepatitis C virus (HCV) is a major cause of liver disease throughout the world. The NS5A and E2 proteins of HCV genotype 1 were reported to inhibit the double-stranded (ds) RNA-dependent protein kinase (PKR), which is involved in the cellular antiviral response induced by interferon (IFN). The response to IFN therapy is quite different between genotypes, with response rates among patients infected with types 2 and 3 that are two-three-fold higher than in patients infected with type 1. Interestingly, a significant percentage of HCV genotype 3-infected patients do not respond to treatment at all. The aim of this paper was to analyse the sequences of fragments of the E2 and NS5A regions from 33 outpatients infected with genotype 3a, including patients that have responded (SVR) or not responded (NR) to treatment. HCV RNA was extracted and amplified with specific primers for the NS5A and E2 regions and the PCR products were then sequenced. The sequences obtained covered amino acids (aa) 636-708 in E2 and in NS5A [including the IFN sensitivity determining region (ISDR), PKR-binding domain and extended V3 region)]. In the E2 and NS5A regions, we did observe aa changes among patients, but these changes were not statistically significant between the SVR and NR groups. In conclusion, our results suggest that the ISDR domain is not predictive of treatment success in patients infected with HCV genotype 3a.publishersversionpublishe

Absence of hepatitis B virus DNA in patients with hepatitis C and non-A-E hepatitis in the State of São Paulo, Brazil

Occult hepatitis B virus (HBV) infection has been reported as cases in which HBV DNA was detected despite the absence of any HBV serological markers or in cases in which anti-HBc antibody was the sole marker. The aim of the present study was to determine, using the polymerase chain reaction (PCR), whether HBV infection occurs in hepatitis C and non-A-E hepatitis patients without serological evidence of hepatitis B infection in São Paulo State. Two different populations were analyzed: 1) non-A-E hepatitis patients, including 12 patients with acute and 50 patients with chronic hepatic disorders without serological evidence of infection with known hepatitis viruses; 2) 43 patients previously diagnosed as hepatitis C with positive results for anti-HCV and HCV RNA. Among hepatitis C patients, anti-HBc was detected in 18.6% of the subjects. Three different sets of primers were employed for HBV DNA detection by nested PCR, covering different HBV genes: C, S and X. HBV-DNA was not detected in any sample, whereas the positive controls did produce signals. The lack of HBV DNA detection with these pairs of primers could be due to a very low viral load or to the presence of mutations in their annealing sites. The latter is unlikely as these primers were screened against an extensive dataset of HBV sequences. The development of more sensitive methods, such as real time PCR, to detect circular covalent closed DNA is necessary in order to evaluate this question since previous studies have shown that cryptic hepatitis B might occur

Elevated Alanine Aminotransferase (alt) In Blood Donors : An Assessment Of The Main Associated Conditions And Its Relationship To The Development Of Hepatitis C

The determination of aminotranferases levels is very useful in the diagnosis of hepatopathies. In recent years, an elevated serum ALT level in blood donors has been associated with an increased risk of post-transfusion hepatitis (PTH). The purpose of the study was to research the factors associated with elevated ALT levels in a cohort of voluntary blood donors and to evaluate the relationship between increased ALT levels and the development of hepatitis C (HCV) infection. 166 volunteer blood donors with elevated ALT at the time of their first donation were studied. All of the donors were questioned about previous hepatopathies, exposure to hepatitis, exposure to chemicals, use of medication or drugs, sexual behaviour, contact with blood or secretions and their intake of alcohol. Every three months, the serum levels of AST, ALT, alkaline phosphatase, gamma glutamyl transpeptidase, cholesterol, triglyceride and glycemia are assessed over a two year follow-up. The serum thyroid hormone levels as well as the presence of auto-antibodies were also measured. Abdominal ultrasound was performed in all patients with persistently elevated ALT or AST levels. A needle biopsy of liver was performed in 9 donors without definite diagnostic after medical investigation. The presence of anti-HCV antibodies in 116 donors were assayed again the first clinical evaluation. At the end of follow-up period (2 years later) 71 donors were tested again for the presence of anti-HCV antibodies. None of donors resulted positive for hepatitis B or hepatitis C markers during the follow-up. Of the 116 donors, 101 (87%) had persistently elevated ALT serum levels during the follow-up. Obesity and alcoholism were the principal conditions related to elevated ALT serum levels in 91/ 101 (90.1%) donors. Hypertriglyceridemia, hypercholesterolemia, hypothyroidism and diabetes mellitus also were associated with increased ALT levels. Only 1/101 (0.9%) had mild chronic active non A-G viral hepatitis and 3/101 (2.9%) had liver biopsy with non-specific reactive hepatitis. The determination of ALT levels was not useful to detect donors infected with HCV at donation in Brazil, including the initial seronegative anti-HCV phase.404219224Aach, R.D., Szmuness, W., Mosley, J.W., Serum alanine aminotransferase of donors in relation to the risk of non-A, non-B hepatitis in recipients (1981) New Engl. J. Med., 304, pp. 989-994Alter, H.J., Purcell, R.H., Holland, P.V., Alling, D.W., Koziol, D.E., Donors transaminase and recipient hepatitis. Impact on blood transfusion services (1981) J. Amer. Med. Ass., 246, pp. 630-634Alter, H.J., The cloning and clinical implications of HGV and HGBV-C (1996) New Engl. J. Med., 334, pp. 1536-1537Alter, H.J., Transfusion transmitted hepatitis C and non-A, non-B, non-C (1994) Vox Sang. 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Med., 107, pp. 137-144Garrow, J.S., (1988) Obesity and Related Diseases, , New York, Churchill LivingstoneGarson, J.A., Tedder, R.S., Briggs, M., Detection of hepatitis C viral sequences in blood donations by "nested" polymerase chain reaction and prediction of infectivity (1990) Lancet, 335, pp. 1419-1422Gonçales Jr., F.L., Boccato, R.S.B.S., Pedro, R.J., Prevalências do HBsAg, do anti-HBc e do anti-HCV na população de candidatos a doadores de sangue do Hemocentro-Campinas (1993) Rev. Inst. Med. Trop. S. Paulo, 35, pp. 45-51Gonçales Jr., F.L., Pedro, R.J., Silva, L.J., Hepatites póstransfusionais na cidade de Campinas, SP, Brasil. II. Presença dos anticorpos anti-HBc e anti-HCV em candidatos a doadores de sangue e ocorrência de hepatites pós-transfusionais pelo virus C nos receptores de sangue ou derivados (1993) Rev. Inst. Med. Trop. S. Paulo, 35, pp. 63-71Gonçales Jr., F.L., Pedro, R.J., Silva, L.J., Hepatites póstransfusionais na cidade de Campinas, SP, Brasil. I. Incidência, agentes etiológicos e aspectos clínico-epidemiológicos da hepatite por virus C (1993) Rev. Inst. Med. Trop. S. Paulo, 35, pp. 53-62Hultcrantz, R., Glaumann, H., Lindberg, G., Nilsson, L.H., Liver investigation in 149 asymptomatic patients with moderately elevated activities of serum aminotransferases (1986) Scand. J. Gastroent., 21, p. 109Ismay, S.L., Thomas, S., Fellows, A., Post-transfusion hepatitis revisited (1995) Med. J. Aust., 163, pp. 74-77Jullien, A.M., Courouce, A.M., Massari, V., Impact of screening donor blood for alanine aminotransferase and antibody to hepatitis B core antigen on the risk of hepatitis C virus transmission (1993) Europ. J. Clin. Microbiol. Infect. Dis., 12, pp. 668-672Kahn, R.A., Johnson, G., Aach, R.D., The distribution of serum alanine aminotransferase levels in a blood donor population (1982) Amer. J. 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(Wash.), 92, pp. 3401-3405Vanderborght, B.O., Reis, A.M., Rouzere, C.D., Prevalence of anti-hepatitis C virus in the blood donor population of Rio de Janeiro (1993) Vox Sang. (Basel), 65, pp. 122-12

Hepatitis B virus genotype E detected in Brazil in an African patient who is a frequent traveler

Genotype E of hepatitis B virus (HBV) has not been described in Brazil and is found mainly in Africa. Genotype A is the most prevalent in Brazil, and genotypes B, C, D, and F have already been reported. We report here an HBV genotype E-infected patient and some characterization of surface (S) protein, DNA polymerase (P) and precore/core (preC/C) coding regions based on the viral genome. The patient is a 31-year-old black man with chronic hepatitis B who was born and raised in Angola. He has been followed by a hepatologist in São Paulo, Brazil, since November 2003, and he is a frequent traveler to Latin America, Africa, and Europe. In 2003, he was diagnosed with HBV infection and started treatment with lamivudine with the later addition of adefovir dipivoxil. No known risk factor was identified. Serologically, he is HBsAg and anti-HBe positive, but HBeAg and anti-HBs negative. DNA sequence analysis of the S/P region confirmed that this patient is infected with genotype E, subtype ayw4. The preC/C region showed G1896A and G1899A mutations but no mutations in the basal core promoter. Nucleotide substitutions common in genotype E were also observed (C1772, T1858 and A1757). Although this is not an autochthonous case and there is no evidence of further spread, the description of this case in Brazil highlights the current risk of viral genotypes spreading with unprecedented speed due to constant travel around the world