[Show abstract][Hide abstract] ABSTRACT: The hepatitis A virus (HAV) HAF-203 strain was isolated from an acute case of HAV infection. The primary isolation of HAF-203 in Brazil and its adaptation to the FRhK-4 cell lineage allowed the production of large amounts of viral particles enabling molecular characterization of the first HAV isolate in Brazil. The aim of our study was to determine the nucleotide sequence of the HAF-203 strain genome, compare it to other HAV genomes and highlight its genetic variability. The complete nucleotide sequence of the HAF-203 strain (7472 nucleotides) was compared to those obtained earlier by others for other HAV isolates. These analyses revealed 19 HAF-specific nucleotide sequence differences with 10 amino acid substitutions. Most of the non-conservative changes were located at VP1, 2C, and 3D genes, but the 3B region was the most variable. The availability of HAF-203 complementary DNA was useful for the production of the recombinant VP1 protein, which is a major determinant of viral infectivity. This recombinant protein was shown by enzyme-linked immunoassay and blotting, to be immunogenic and resemble the native protein, therefore suggesting its value as a reagent for incorporation into diagnostic tests.
Memórias do Instituto Oswaldo Cruz 12/2006; 101(7):759-66. · 1.36 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In order to investigate the hepatitis C virus (HCV) infection prevalence and risk factors in hemophiliacs in Central Brazil, 90 patients were interviewed and serum samples tested for HCV RNA and anti-HCV antibodies. An overall prevalence of 63.3% (CI 95%: 53.0-72.7) was found. Multivariate analysis of risk factors showed that number of blood transfusions was significantly associated with this infection. Most hemophiliacs received locally produced cryoprecipitate. All infected patients were transfused before the screening of blood units for anti-HCV. However, hemophiliacs who received exclusively screened cryoprecipitate were HCV negative. It confirms the expected decline in transfusion-acquired hepatitis C.
Memórias do Instituto Oswaldo Cruz (ISSN: 1678-8060) Vol 97 Num 5. 01/2002;
[Show abstract][Hide abstract] ABSTRACT: An hemodialysis population in Central Brazil was screened by polymerase chain reaction (PCR) and serological methods to assess the prevalence of hepatitis C virus (HCV) infection and to investigate associated risk factors. All hemodialysis patients (n=428) were interviewed in eight dialysis units in Goiânia city. Blood samples were collected and serum samples screened for anti-HCV antibodies by an enzyme-linked immunosorbent assay (ELISA). Positive samples were retested for confirmation with a line immunoassay (LIA). All samples were also tested for HCV RNA by the PCR. An overall prevalence of 46.7% (CI 95%: 42-51.5) was found, ranging from 20.7% (CI 95%: 8.8-38.1) to 90.4% (CI 95%: 79.9-96.4) depending on the dialysis unit. Of the 428 patients, 185 were found to be seropositive by ELISA, and 167 were confirmed positive by LIA, resulting in an anti-HCV prevalence of 39%. A total of 131 patients were HCV RNA-positive. HCV viremia was present in 63.5% of the anti-HCV-positive patients and in 10.3% of the anti-HCV-negative patients. Univariate analysis of risk factors showed that the number of previous blood transfusions, transfusion of blood before mandatory screening for anti-HCV, length of time on hemodialysis, and treatment in multiple units were associated with HCV positivity. However, multivariate analysis revealed that blood transfusion before screening for anti-HCV and length of time on hemodialysis were significantly associated with HCV infection in this population. These data suggest that nosocomial transmission may play a role in the spread of HCV in the dialysis units studied. In addition to anti-HCV screening, HCV RNA detection is necessary for the diagnosis of HCV infection in hemodialysis patients.
Memórias do Instituto Oswaldo Cruz 09/2001; 96(6):765-9. · 1.36 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Eight cases of de novo hepatitis C virus (HCV) infection in a haemodialysis unit in Rio de Janeiro, Brazil, were retrospectively studied. HCV viraemia was demonstrated by RT nested PCR in seven of the seroconverters. Genotyping showed that six patients were infected with a genotype 1b strain and one with a genotype 1a strain. A phylogenetic analysis of nucleotide sequences of the HCV core region revealed that five of the six 1b isolates form a separate cluster when compared with other 38 HCV 1b core sequences randomly chosen from the GenBank. The revealed sequence similarities indicated the nosocomial spread of a single HCV strain within the unit. To investigate whether the patients infected with the same viral isolate display similar patterns of antibody response to individual proteins, serial serum samples were examined. A line immunoassay for qualitative and semi-quantitative determination of specific antibodies against recombinant and synthetic HCV antigens was employed. Despite infection with the same virus strain, the patients sera demonstrated different patterns of reactivity against individual structural and nonstructural HCV proteins immediately after seroconversion. For each patient, however, antibody responses remained mostly stable throughout the follow-up of 8 to 24 months.
Journal of Medical Virology 05/2001; 63(4):265-70. · 2.37 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: During an outbreak of hepatitis B virus (HBV) infection in a hemodialysis unit, patients were assessed for serological viral markers and vaccination status. HBV infection was identified in 26 patients. Twenty of these were positive for hepatitis B surface antigen (HBsAg), and 6 were negative for HBsAg but positive for IgM antibody to hepatitis B core (anti-HBc) and HBV DNA. The primary source of infection was not clearly identified, although 2 patients were suspected to be the index cases. A multiple logistic regression analysis revealed low anti-HBs titers and vaccination status to be independently associated with the risk of acquiring HBV infection. Both the high prevalence of HBV infection (31%) detected in this unit and the low vaccine response (53%) observed reinforce the importance of universal and preventive measures in controlling HBV infection. The detection of HBV DNA in HBsAg-negative/IgM anti-HBc-positive patients emphasizes the value of anti-HBc testing in the routine screening of HBV in hemodialysis units.
[Show abstract][Hide abstract] ABSTRACT: A retrospective study on the prevalence of hepatitis E virus (HEV) infection was conducted in selected populations in Rio de Janeiro, Brazil. A total of 1,115 subjects were tested including 146 patients with acute Non-A Non-B Non-C (NANBNC) viral hepatitis, 65 hemodialysis patients, 93 blood donors, 102 intravenous drug users (IVDUs), 304 pregnant women, 145 individuals living in the rural area and 260 individuals living in the urban area. In order to characterize a favorable epidemiological set for enterically transmitted infection in the studied populations we also evaluated the prevalence of anti-HAV IgG (hepatitis A virus) antibodies. Specific antibodies to HEV (anti-HEV IgG) were detected by a commercial EIA and specific antibodies to HAV (anti-HAV IgG) were detected using a competitive "in house" EIA. We found a high prevalence of anti-HAV IgG in these populations, that could indicate some risk for infections transmitted via the fecal-oral route. The anti-HEV IgG prevalence among the different groups were: 2.1% in patients with acute NANBNC viral hepatitis, 6.2% in hemodialysis patients, 4.3% in blood donors, 11.8% in IVDUs, 1% in pregnant women, and 2.1% in individuals form the rural area. Among individuals living in the urban area we did not find a single positive serum sample. Our results demonstrated the presence of anti-HEV IgG in almost all studied populations; however, further studies are necessary to establish the real situation of HEV epidemiology in Rio de Janeiro, Brazil.
Memórias do Instituto Oswaldo Cruz 02/2001; 96(1):25-9. · 1.36 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The prevalence of antibodies directed against the enterically transmitted hepatitis A virus (HAV) was measured in 2 groups of people living in Rio de Janeiro, Brazil. Of 1,056 health care workers (HCWs), 778 (73.7%) were anti-HAV positive. A high prevalence of anti-HAV antibodies (85.7%) was also found among 274 voluntary blood donors (BDs). TT virus (TTV) is a DNA virus that has been found in the sera of patients with post-transfusion hepatitis of unknown etiology. Occurrence of virus shedding suggests that the fecal-oral route may be an important mode of TTV transmission, particularly in the developing world. The presence of TTV DNA was analyzed by PCR in the sera of 191 HCWs and 151 BDs. TTV was detected in 65.4% of HCWs and 79.5% of BDs. In both groups, a family income of < US$400 per month and a level of education of < 11 y of schooling were found to be risk factors for HAV infection. Furthermore, a low family income was associated with TTV viremia in the HCW group. However, the presence of TTV DNA was associated with neither low level of education nor anti-HAV positivity.
[Show abstract][Hide abstract] ABSTRACT: The objective of this investigation was to study the prevalence of serological markers of hepatitis B and possible risk factors for this disease in a sample of 404 people who attended a Testing and Couseling Center for HIV in the city of Ribeirão Preto, São Paulo State, Brazil. The overall prevalence of serologic hepatitis B markers was 14.6%, equal to that obtained for anti-HBc. HBsAg and anti-HBc IgM showed prevalences of 1%. After adjustment using logistic regression, hepatitis B markers showed association with the following variables: age, place of residence, use of injectable drugs and positivity to anti-HIV. The overall prevalence of human immunodeficiency virus infection was 6.9%. Hepatitis B markers were detected in 55.6% among intravenous drug users and in 42.9% among those who tested positive for HIV, confirming literature findings which indicates high levels of infection in these specific population groups.
Revista da Sociedade Brasileira de Medicina Tropical 01/2001; 34(1):53-9. · 0.93 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A method for genotyping hepatitis B virus (HBV) strains, based on restriction fragment length polymorphism (RFLP) analysis of four different amplified fragments of the HBV genome, was used to investigate nosocomial infections that occurred in two Brazilian hemodialysis centers. Viral isolates from hepatitis B surface antigen (HBsAg)-positive serum samples from 27 hemodialysis patients and 39 HBV-positive unrelated control patients were grouped according to their RFLP patterns. Strains isolated from the control patients were divided into nine RFLP patterns: A1, A2, A3 (genotype A), D1, D2, D3, D4 (genotype D), F1, and F2 (genotype F). In hemodialysis unit A (Rio de Janeiro), 14 HBV isolates were grouped into five different RFLP patterns: A1, A2, A3, D3, and D4. Pattern A2, present at a relatively low prevalence (18%) in the control group, was observed in the majority (53%) of the hemodialysis patients. Notably, all five patients who seroconverted to HBsAg positivity in 1995 carried the strain A2. In hemodialysis unit B (state of São Paulo), where an outbreak of HBV infection occurred in 1996-1997, RFLP analysis showed that all 13 patients who seroconverted were infected with HBV isolates of genotype D. Coinfection with strain A1 was detected in seven of them. The results demonstrate the value of RFLP analysis in establishing common sources of infection in hemodialysis centers.
European Journal of Clinical Microbiology 08/2000; 19(7):531-7. · 3.02 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A survey was carried out in 2 drug use treatment centres (TCs) in Rio de Janeiro, Brazil, to assess risk behaviours, HIV infection and other sexually transmitted infections/blood-borne infections (STIs/BBIs). Two hundred and twenty-five drug users (195 males and 30 females) were interviewed and clinically examined, and their blood and urine were tested for STIs/BBIs. Prevalences (%) for these infections were as follows--HIV: 0.9, hepatitis B virus (HBV): 14.7, hepatitis C virus (HCV): 5.8, syphilis: 5.3, gonorrhoea/chlamydia (CT/NG): 4.7. In bivariate analyses CT/NG infection was associated with younger age (P=0.003); current genitourinary symptoms (odds ratio [OR]=6.2) and a mainly illegal source of income (OR=9.1). Hepatitis C infection was associated with a history of ever having injected any drug (OR=19.6), and with each one of the injected drugs. After multiple logistic regression, lower educational level (adjusted odds ratio [AOR]=3.70) and 'ever having injected drugs' (AOR=3.69) remained as independent risk factors for hepatitis B infection. In conclusion, TCs must implement programmes directed towards the prevention of STIs/BBIs.
International Journal of STD & AIDS 07/2000; 11(6):383-92. · 1.00 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To study some of the epidemiological aspects of hepatitis B in a non-representative sample of patients seen in health care clinics.
The study population comprised 632 patients who were seen at health care clinics in the city of Ribeirão Preto, Brazil, for the purpose of blood testing, regardless the reason. After signing a written consent, an additional amount of blood was drawn from the same venous puncture site used to collect the original sample for the testing assigned to the patient at the health care clinic. A questionnaire was applied to each participant, looking for the presence of risk factors for hepatitis B. The blood samples were tested for HBV markers, using immunoenzimatic techniques.
The prevalences of HBsAg and anti-HBcAg were 0.3% and 13.9%, respectively. By a logistic regression model, the following variables were significantly associated with the infection: age, time of residency in the city (higher risk among those living for a period less than one year), past history of hepatitis, incarceration and sexual behavior (higher risk among homosexual and bisexual males).
The growing difficulties in obtaining blood samples from a representative group of patients, as done in classic surveys, make it necessary to look for alternative methodologies which can provide information concerning the presence of infectious agents in a community. Though the results cannot be generalized to the population as a whole, the methodology used conveyed some knowledge regarding the circulation of hepatitis B virus. In addition, it makes much easier to obtain agreement from the participants, since it does not add any invasive procedure. Despite the limitations, this methodology may be helpful in epidemiological surveillance of infectious agents known as producing asymptomatic infections in much of the population.
Revista de Saúde Pública 07/2000; 34(3):286-91. · 1.07 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: There are no data concerning the genotypic analysis of hepatitis B virus (HBV) infection in Brazilian dialysis centers. Serum samples from all hemodialysis patients (n = 282) in Goiânia City, Central Brazil, were tested for hepatitis B surface antigen (HBsAg). An overall prevalence of 12.0% was found, ranging from 0 to 33.3% depending on dialysis centers. Positive samples (n = 34) were submitted to serological subtyping by monoclonal ELISA and HBV DNA detection by polymerase chain reaction (PCR). Among 30 PCR-positive samples, 26 were genotyped by use of the line probe assay technology (INNO-LiPA HBV, Innogenetics, Gent, Belgium). HBV genotypes A (50. 0%) and D (46.2%) were the most frequently found whereas genotype F (3.8%) was rare in this population. Serological subtypes adw2 (44. 1%) and ayw3 (41.2%) were dominant. By contrast, adw4 and ayw2 were found at a low frequency (2.9%). A correlation was observed in the distribution of genotypes and subtypes by dialysis center. Genotype D and subtype ayw3 were predominant in 2 hemodialysis centers whereas genotype A and subtype adw2 were predominant in the others. The findings of high HBsAg prevalence rates restricted to certain dialysis centers and the data obtained through genotyping and serological subtyping suggest HBV nosocomial transmission in these hemodialysis centers.
[Show abstract][Hide abstract] ABSTRACT: Viral hepatitis constitutes a major health issue, with high prevalence among injecting drug users (IDUs). The present study assessed the prevalence and risk determinants for hepatitis B, C and D viruses (HBV, HCV and HDV) infections among 102 IDUs from Rio de Janeiro, Brazil. Serological markers and HCV-RNA were detected by enzyme immunoassay and nested PCR, respectively. HCV genotyping was determined by restriction fragment length polymorphism analysis (RFLP). HBsAg, anti-HBc and anti-HBs were found in 7.8, 55.8 and 24. 7% of IDUs, respectively. In the final logistic regression, HBV infection was independently associated with male homosexual intercourse within the last 5 years (odds ratio (OR) 3.1; 95% confidence interval (CI) 1.1-8.8). No subject presented anti-delta (anti-HD). Anti-HCV was detected in 69.6% of subjects, and was found to be independently associated with needle sharing in the last 6 months (OR 3.4; 95% CI 1.3-9.2) and with longer duration of iv drug use (OR 3.1; 95% CI 1.1-8.7). These data demonstrate that this population is at high risk for both HBV and HCV infection. Among IDUs from Rio de Janeiro, unprotected sexual intercourse seems to be more closely associated with HBV infection, whereas HCV is positively correlated with high risk injecting behavior. Comprehensive public health interventions targeting this population and their sexual partners must be encouraged.
Brazilian Journal of Medical and Biological Research 10/1999; 32(9):1107-14. · 1.14 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Hepatitis C virus (HCV) infection is widespread and responsible for more than 60% of chronic hepatitis cases. HCV presents a genetic variability which has led to viral classification into at least 6 genotypes and a series of subtypes. These variants present characteristic geographical distribution, but their association with different responses to treatment with interferon and severity of disease still remains controversial. The aim of this study was to investigate the patterns of distribution of HCV genotypes among different exposure categories in Brazil. Two hundred and fifty anti-HCV positive samples were submitted to HCV-RNA detection by RT-PCR and their genotype was determined by restriction fragment length polymorphism (RFLP) analysis. In addition, the genotype/subtype of 60 samples was also determined by a reverse hybridization assay. HCV 1 was the most prevalent (72.0%), followed by type 3 (25.3%), HCV 2 (2.0%) and HCV 4 (0.7%). The HCV genotype distribution varied among the different exposure categories, with HCV 1 being more frequent among blood donors, hemophiliacs and hemodialysis patients. A high frequency of HCV 3 was observed in cirrhotic patients, blood donors from the South of Brazil and injecting drug users (IDUs). The general distribution of the HCV genotype in Brazil is similar to that in other regions of the world.
Brazilian Journal of Medical and Biological Research 04/1999; 32(3):279-82. · 1.14 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Hepatitis viruses belong to different families and have in common a striking hepatotropism and restrictions for propagation in cell culture. The transmissibility of hepatitis is in great part limited to non-human primates. Enterically transmitted hepatitis viruses (hepatitis A virus and hepatitis E virus) can induce hepatitis in a number of Old World and New World monkey species, while the host range of non-human primates susceptible to hepatitis viruses transmitted by the parenteral route (hepatitis B virus, hepatitis C virus and hepatitis delta virus) is restricted to few species of Old World monkeys, especially the chimpanzee. Experimental studies on non-human primates have provided an invaluable source of information regarding the biology and pathogenesis of these viruses, and represent a still indispensable tool for vaccine and drug testing.
Brazilian Journal of Medical and Biological Research 09/1998; 31(8):1035-48. · 1.14 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Hepatitis C virus (HCV) is considered to be the major agent of viral hepatitis, resulting in chronic liver disease, cirrhosis, and hepatocellular carcinoma. Eleven HCV types, including at least 80 subtypes, have already been described and shown to have different geographical distribution. Several reports have suggested that HCV genotypes have distinct clinical outcomes with regard to disease severity and response to alfa-interferon. Differences in serological reactivity between HCV genotypes have also been reported. In addition, vaccines against multiples HCV genotypes will be necessary and, therefore, mapping the distribution of viral genotypes is warranted. However, little is known about the genotypic analysis of HCV in Latin America. In Brazil, previous studies were carried out in Rio de Janeiro, Sao Paulo (southeastern region), and Porto Alegre (south region). Given the large size of the country, its mixture of cultures and races, the diversity in climatological regions and social conditions, and the limited amount of data concerning HCV genotypes in other Brazilian regions, we decided to investigate the distribution of HCV types and subtypes in blood donors from three different geographical regions of Brazil.
Memórias do Instituto Oswaldo Cruz 05/1998; · 1.36 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Since the identification of the new human virus, GB virus C (GBV-C)/hepatitis G-virus (HGV), in 1995/1996, reverse transcription polymerase chain reaction remained the sole available diagnostic tool for GBV-C/HGV infection. Recently, a serologic test based on the detection of antibodies to the putative envelope protein 2 (anti-E2) has been introduced. We used this assay for a seroepidemiological survey including 3,314 healthy individuals from different parts of the world, 123 patients from Germany who were suspected to have an increased risk of acquiring GBV-C/HGV infection, 128 multiple organ donors, and 90 GBV-C/HGV RNA positive persons. In European countries, anti-E2 seropositivity ranged from 10.9% (Germany) to 15.3% (Austria). In South Africa (20.3%) and Brazil (19.5%), even higher anti-E2 prevalence rates were recorded. In Asian countries like Bhutan (3.9%), Malaysia (6.3%), and the Philippines (2.7%), anti-E2 positivity was significantly lower. GBV-C/HGV anti-E2 prevalence in potential "risk groups," i.e., patients on hemodialysis and renal transplant recipients, did not vary significantly from anti-E2 seroprevalence in German blood donors. Anti-E2 and GBV-C/HGV RNA were found to be mutually exclusive, confirming the notion that anti-E2 has to be considered as a marker of past infection.
Journal of Medical Virology 03/1998; 54(2):103-6. · 2.37 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The recently discovered hepatitis G virus (HGV) belongs, as hepatitis C virus (HCV), to the Flaviviridae family. HGV has been isolated from the serum of patients with non A-E hepatitis. However, the association of HGV with hepatitis is uncertain.
To determine the HGV prevalence in blood donors and in patients with liver disease and to evaluate a possible correlation between HGV infection and liver disease.
Sera from a total of 113 consecutive patients with chronic liver disease were submitted to a series of liver enzymes and function tests and analyzed for the presence of HBsAg, anti-HBs, anti-HBc, anti-HCV, HCV RNA and HGV RNA. Prevalence of HGV RNA was determined in a group of 87 blood donors.
Nine (10%) sera from blood donors and 15 (13%) sera from patients with chronic liver disease were HGV RNA positive. Some 28 (25%) patients were HCV RNA positive, with genotypes 1a, 1b and 3 present in 10, 12 and 5 patients, respectively. A total of 20 (18%) patients were HBsAg carriers. Five (4%) patients were double infected (one with HBV + HCV, one with HBV + HGV and three with HCV + HGV).
The proportion (10%) of HGV-infected blood donors was very high when compared with other countries. The results did not allow to establish HGV as an etiologic agent for chronic liver disease. The parenteral route was the presumed means of HGV transmission for only one-third of the patients.
Clinical and Diagnostic Virology 02/1998; 9(1):1-7.
[Show abstract][Hide abstract] ABSTRACT: The age-specific prevalence of antibodies to hepatitis A virus (anti-HAV) was determined in two different population groups with low socio-economic status from Rio de Janeiro city, Brazil, whose serum samples were collected 17 years apart (Population 1, 1978; Population 2, 1995). In Population 2, analysis of the anti-HAV prevalence was also carried out with respect to environmental factors. Population 1 was composed of 520 stored sera collected from the umbilical cord of term neonates and children aged 1 month to 6 years. In population 2, 720 serum samples were collected from children and adolescents with ages ranging from 1 to 23 years. The overall prevalence rate of anti-HAV in Population 1 and Population 2 was 65.6% and 32.1%, respectively. In Population 1, the anti-HAV prevalence reached 88% at the age of 3, while in Population 2, it increased from 4.5% in children under the age of 3 to 66% in the group of adolescents over the age of 14. The low exposure to HAV infection in younger children from Population 2 could be a result of improved environmental hygiene and sanitation, as demonstrated by the presence of piped water, waste and sewage disposal systems in most houses from this population group. These findings indicate a possible change in the prevalence of hepatitis A in Rio de Janeiro.
Memórias do Instituto Oswaldo Cruz 01/1998; 93(1):1-5. · 1.36 Impact Factor