The emergence of YMDD mutants precedes biochemical flare by 19 weeks in lamivudine-treated chronic hepatitis B patients: an opportunity for therapy reevaluation

Given the loss of therapeutic efficacy associated with the development of resistance to lamivudine (LMV) and the availability of new alternative treatments for chronic hepatitis B patients, early detection of viral genotypic resistance could allow the clinician to consider therapy modification before viral breakthrough and biochemical relapse occur. To this end, 28 LMV-treated patients (44 ± 12 years; 24 men), on their first therapy schedule, were monitored monthly at four Brazilian centers for the emergence of drug resistance using the reverse hybridization-based INNO-LiPA HBV DR assay and occasionally sequencing (two cases). Positive viral responses (HBV DNA clearance) after 6, 12, and 18 months of therapy were achieved by 57, 68, and 53 percent of patients, while biochemical responses (serum alanine aminotransferase normalization) were observed in 82, 82, and 53 percent of cases. All viral breakthrough cases (N = 8) were related to the emergence of YMDD variants observed in 7, 21, and 35 percent of patients at 6, 12, and 18 months, respectively. The emergence of these variants was not associated with viral genotype, HBeAg expression status, or pretreatment serum alanine aminotransferase levels. The detection of resistance-associated mutations was observed before the corresponding biochemical flare (41 ± 14 and 60 ± 15 weeks) in the same individuals. Then, if highly sensitive LMV drug resistance testing is carried out at frequent and regular intervals, the relatively long period (19 ± 2 weeks) between the emergence of viral resistance and the onset of biochemical relapse can provide clinicians with ample time to re-evaluate drug therapy.

The emergence of YMDD mutants precedes biochemical flare by 19 weeks in lamivudine-treated chronic hepatitis B patients: an opportunity for therapy reevaluation.

Given the loss of therapeutic efficacy associated with the development of resistance to lamivudine (LMV) and the availability of new alternative treatments for chronic hepatitis B patients, early detection of viral genotypic resistance could allow the clinician to consider therapy modification before viral breakthrough and biochemical relapse occur. To this end, 28 LMV-treated patients (44 +/- 12 years; 24 men), on their first therapy schedule, were monitored monthly at four Brazilian centers for the emergence of drug resistance using the reverse hybridization-based INNO-LiPA HBV DR assay and occasionally sequencing (two cases). Positive viral responses (HBV DNA clearance) after 6, 12, and 18 months of therapy were achieved by 57, 68, and 53% of patients, while biochemical responses (serum alanine aminotransferase normalization) were observed in 82, 82, and 53% of cases. All viral breakthrough cases (N = 8) were related to the emergence of YMDD variants observed in 7, 21, and 35% of patients at 6, 12, and 18 months, respectively. The emergence of these variants was not associated with viral genotype, HBeAg expression status, or pretreatment serum alanine aminotransferase levels. The detection of resistance-associated mutations was observed before the corresponding biochemical flare (41 +/- 14 and 60 +/- 15 weeks) in the same individuals. Then, if highly sensitive LMV drug resistance testing is carried out at frequent and regular intervals, the relatively long period (19 +/- 2 weeks) between the emergence of viral resistance and the onset of biochemical relapse can provide clinicians with ample time to re-evaluate drug therapy.

Hepatitis C prevalence and risk factors in hemodialysis patients in Central Brazil: a survey by polymerase chain reaction and serological methods.

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.

A recombinant peptide antigen line immunoassay optimized for the confirmation of Chagas' disease.

BACKGROUND: The transfusion of contaminated blood has become the major route of transmission for Chagas' disease in Brazil. Current screening tests are insensitive and yield conflicting results, while confirmatory assays do not exist. A line immunoassay (INNO-LIA Chagas Ab [INNO-LIA]) combining relevant, immunodominant recombinant and synthetic antigens on a single nylon membrane strip was evaluated for the serologic confirmation of Chagas' disease. STUDY DESIGN AND METHODS: Sera from 1062 patients and healthy residents of four Brazilian regions endemic for Chagas' disease were used for test optimization. The established confirmation algorithm was evaluated with an independent set of positive (n = 75) and negative (n = 148) samples. RESULTS: In the optimization phase, without an established comparative gold standard, the results with the INNO-LIA were compared with those obtained in four other screening assays. In the validation phase, the INNO-LIA showed a sensitivity of 100 percent (95% CI, 95.21-100) and a specificity of 99.32 percent (95% CI, 96.29-99.98) for well-characterized sera. Moreover, its specificity reached 100 percent with a set of 40 sera obtained from patients with documented leishmaniasis. The interpretation criteria defined in this study indicated that the INNO-LIA accurately detected the presence of antibodies to various specific antigens of Trypanosoma cruzi. CONCLUSION: The INNO-LIA Chagas Ab assay may become the first commercial assay to reliably confirm the presence of antibodies to T. cruzi.

Hepatitis B virus infection profile in central Brazilian hemodialysis population.

Hepatitis B has proved to be a major health hazard in hemodialysis patients. In order to investigate the hepatitis B virus (HBV) infection profile in the hemodialysis population of Goiânia city--Central Brazil, all dialysis patients (N = 282) were studied. The prevalence of any HBV marker (HBsAg, anti-HBs, and anti-HBc) was 56.7% (95% CI: 51.1-62.7), ranging from 33.3% to 77.7% depending on dialysis unit. HBV-DNA was detected in 67.6% and 88.2% of the HBsAg-positive serum samples, in 91.3% and 100% of the HBsAg/HBeAg-positive samples, and in 18.2% and 63.6% of the HBsAg/anti-HBe-reactive sera by hybridization and PCR, respectively. The length of time on hemodialysis was significantly associated with HBV seropositivity. Only 10% of the patients reported received hepatitis B vaccination. The findings of a high HBV infection prevalence in this population and the increased risk for HBV infection on long-term hemodialysis suggest the environmental transmission, emphasizing the urgent need to evaluate strategies of control and prevention followed in these units.

Short report: prevalence of hepatitis C viral antibody among Brazilian children, adolescents, and street youths.

The prevalence of hepatitis C antibodies (anti-HCV) was investigated in 1,378 central Brazilian children, students, and street youths (homeless adolescents without family links or adolescents working in poorly paid activities). Sera were tested with a second generation enzyme-linked immunosorbent assay, and positive samples were retested by a confirmatory assay (line immunoassay). All children attending day care centers were anti-HCV negative. Only one (0.2%) adolescent was positive in the student group. However, higher positivity was found in street youths; four (1.0%) living at home and three (3.0%) living in the streets and anti-HCV antibodies. Among these, the prevalence of anti-HCV increased significantly with age from 0% in the 9-12-year-old group to 6.9% in the 17-20-year-old group. Risk factors including blood transfusion, tattooing, intravenous drug use, and sexual intercourse with multiple partners were significantly associated with the presence of anti-HCV in street youths. These results indicate that apparently healthy children and adolescents attending day care centers or primary schools in central Brazil have a low exposure to HCV infection, but street youths in the same area are at risk for infection with this virus.

High prevalence of hepatitis C infection among brazilian hemodialysis patients in Rio de Janeiro a one year follow-up study

Aproxidamente 400 pacientes de hemodialise tratados em 5 diferentes unidades no Rio de Janeiro foram acompanhados durante 1 ano para presenca de marcadores virais de hepatite B e C. Durante o mesmo periodo, amostras foram tambem de 35 pacientes ambulatoriais de dialise peritonial continua (CAPD) e de 242 funcionarios das unidades. Dependendo da unidade em estudo foram detectadas prevalencias de anti-HCV variando de 47 por cento a 82 por cento (media 65 por cento ). Pacientes de prevalencia de anti-HCV em funcionarios foi de 2,9 por cento . Observamos uma taxa de ataque de hepatite C de 11,5 por cento por ano na populacao paciente de hemodialise anti-HCV negativo. Uma media de 9,4 por cento de pacientes de hemodialise eram portadores cronicos do virus da hepatite B (VHB) (taxa de 1.8 por cento a 20.4 por cento ), enquanto 48.9 por cento apresentaram marcadores de infeccao passada de HBV. A taxa de ataque de HBV foi de 4.5 por cento por ano (taxa de 0 por cento a 6 por cento ). Esses resultados indicam uma alarmante prevalencia alta anti-HCV em pacientes de hemodialise dessa regiao estudada.

High prevalence of hepatitis C infection among Brazilian hemodialysis patients in Rio de Janeiro: a one-year follow-up study.

Nearly 400 hemodialysis patients treated at 5 different hemodialysis units in Rio de Janeiro were tested for one year for the presence of hepatitis C and B markers. During the same period, samples were also obtained from 35 continuous ambulatory peritoneal dialysis (CAPD) patients and from 242 health care workers. Depending on the hemodialysis unit studied, anti-HCV prevalence rates ranging from 47% to 82% (mean 65%) were detected. CAPD patients showed a lower prevalence of 17%. The prevalence of antibodies against hepatitis C virus (anti-HCV) among health care workers was 2.9%. We observed a hepatitis C attack rate of 11.5% per year in the anti-HCV-negative hemodialysis patient population. An average of 9.4% of the hemodialysis patients were chronic carriers of hepatitis B virus (HBV) (range 1.8% - 20.4%), while 48.9% showed markers of previous HBV infection. The HBV attack rate was 4.5% per year (range 0% - 6%). These results indicate an alarming high prevalence of anti-HCV among hemodialysis patients of this studied region.

Hepatitis B and C prevalences among blood donors in the south region of Brazil

The prevalence of hepatitis B and C infection has been determined in a seroepidemiological survey among blood donors from the south of Brazil (Florianópolis, State of Santa Catarina). These markers has also been correlated with the levels of alanine aminotransferase (ALT), a surrogate marker to prevent post-transfusion hepatitis. Sera from 5000 donors were randomly collected in the period of April to November 1991. The prevalences of HBsAg, anti-HBs and anti-HBc were respectively 0.78, 7.02 and 13.98. The anti-HCV prevalence after confirmation testing with line immunoassay (LIA), was 1.14. Normal values of ALT ( < = 32 U/ml) were found in 59.78, values slightly above the mean (ALT between 32-70 U/ml) in 37.74 and high values of ALT ( > = 70 U/ml) in 2.48. The positivity of anti-HCV antibodies increased with the elevation of ALT levels. This correlation was not observed in relation to HBsAg. There exists a diversity in the recognition of HCV epitopes among HCV positive donors. Via the confirmation test used, we could observe that 94.7 of donors recognize the structural core antigen. Besides that, we observed that 5.26 of the HCV reactive sera recognized only epitopes located in the NS4 and/or NS5 region, indicating the importance of these epitopes for the improvement of assays.

Anti-HCV related to HCV PCR and risck factors analysis in a blood donor population of central Brazil

Ainda sao raros os casos de infeccao por hepatite C (HCV)na regiao central do Brasil. Neste estudo, 2.350 doadores voluntarios de sangue foram avaliados, resultando em prevalencias para o anti-HCV de 2,2 (por cento), pelo ELISA de segunda geracao, e de 1,4 por cento, apos o ensaio confirmatorio "line immunoassay". Anticorpos contra os antigenos "core", NS4 e NS5 do HCV foi observada em 76,6 (por cento) dos doadores anti-HCV positivos. A positividade da reacao em cadeia da polimerase (PCR) mostrou-se relacionada a reatividade aos diferentes antigenos do HCV no "line immunoassay". A maioria dos doadores positivos tiveram historia previa de exposicao parenteral. A combinacao de ALT>50 UI/1 e positividade ao anti-HBc parece nao ser eficaz como marcadores indiretos para a infeccao pelo HCV, entretanto a dosagem do ALT e a deteccao de anti-HCV sao indicadas na triagem de doadores de sangue brasileiros.

Anti-HCV related to HCV PCR and risk factors analysis in a blood donor population of central Brazil.

Data concerning HCV infection in Central Brazil are rare. Upon testing 2,350 voluntary blood donors from this region, we found anti-HCV prevalence rates of 2.2% by a second generation ELISA and 1.4% after confirmation by a line immunoassay. Antibodies against core, NS4, and NS5 antigens of HCV were detected in 81.8%, 72.7%, and 57.5%, respectively, of the positive samples in the line immunoassay. HCV viremia was present in 76.6% of the anti-HCV-positive blood donors. A relation was observed between PCR positivity and serum reactivity in recognizing different HCV antigens in the line immunoassay. The majority of the positive donors had history of previous parenteral exposure. While the combination of ALT > 50 IU/l and anti-HBc positivity do not appear to be good surrogate markers for HCV infection, the use of both ALT anti-HCV tests is indicated in the screening of Brazilian blood donors.

Hepatitis B and C prevalences among blood donors in the south region of Brazil.

The prevalence of hepatitis B and C infection has been determined in a seroepidemiological survey among blood donors from the south of Brazil (Florianópolis, State of Santa Catarina). These markers has also been correlated with the levels of alanine aminotransferase (ALT), a surrogate marker to prevent post-transfusion hepatitis. Sera from 5000 donors were randomly collected in the period of April to November 1991. The prevalences of HBsAg, anti-HBs and anti-HBc were respectively 0.78%, 7.02% and 13.98%. The anti-HCV prevalence after confirmation testing with line immunoassay (LIA), was 1.14%. Normal values of ALT ( < = 32 U/ml) were found in 59.78%, values slightly above the mean (ALT between 32-70 U/ml) in 37.74% and high values of ALT ( > = 70 U/ml) in 2.48%. The positivity of anti-HCV antibodies increased with the elevation of ALT levels. This correlation was not observed in relation to HBsAg. There exists a diversity in the recognition of HCV epitopes among HCV positive donors. Via the confirmation test used, we could observe that 94.7% of donors recognize the structural core antigen. Besides that, we observed that 5.26% of the HCV reactive sera recognized only epitopes located in the NS4 and/or NS5 region, indicating the importance of these epitopes for the improvement of assays.

Antibodies against non-structural c100/3 and structural core antigen of hepatitis C virus (HCV) in hemodialysis patients.

Two groups of patients undergoing hemodialysis (HD) maintenance were evaluated for their antibody response to non-structural c100/3 protein and structural core protein of hepatitis C virus (HCV). Forty-six patients (Group 1) never presented liver abnormalities during HD treatment, while 52 patients (Group 2) had either current or prior liver enzyme elevations. Prevalence rates of 32.6% and 41.3% were found for anti-c100/3 and anti-HCV core antibodies, respectively, in patients with silent infections (Group 1). The rate of anti-c100/3 in patients of Group 2 was 71.15% and reached 86.5% for anti-HCV core antibodies. The recognition of anti-c100/3 and anti-core antibodies was significantly higher in Group 2 than in Group 1. A line immunoassay composed of structural and non-structural peptides was used as a confirmation assay. HBV infection, measured by the presence of anti-HBc antibodies, was observed in 39.8% of the patients. Six were HBsAg chronic carriers and 13 had naturally acquired anti-HBs antibodies. The duration of HD treatment was correlated with anti-HCV positivity. A high prevalence of 96.7% (Group 2) was found in patients who underwent more than 5 years of treatment. Our results suggest that anti-HCV core ELISA is more accurate for detecting HCV infection than anti-c100/3. Although the risk associated with the duration of HD treatment and blood transfusion was high, additional factors such as a significant non-transfusional spread of HCV seems to play a role as well. The identification of infective patients by more sensitive methods for HCV genome detection should help to control the transmission of HCV in the unit under study.

Prevalence of anti-hepatitis C virus in the blood donor population of Rio de Janeiro.

The prevalence of hepatitis C virus (HCV) antibodies in 2,557 asymptomatic volunteer Brazilian blood donors is reported. Using the line immunoassay (Inno-LIA) as a confirmatory test on ELISA anti-HCV-positive reacting sera, a prevalence rate of 2.7% for anti-HCV positivity was found. By comparison, prevalences of 1.6% for hepatitis B surface antigen, 0.9% for Treponema pallidum, 0.4% for human immunodeficiency virus and 0.04% for Trypanosoma cruzi were observed. Only 57% of the HCV-positive donors had elevated alanine aminotransferase (ALT) levels. Using previous criteria, based on surrogate markers (ALT > or = 50 IU/l and for anti-hepatitis B core antibody), for HCV infection at that time, only 25% of the HCV-positive donations would have been eliminated. In view of the high prevalence of anti-HCV reactivity among the Brazilian blood donor population and the poor reliability of surrogate markers, it is recommended that routine screening for anti-HCV in Brazilian blood donors is introduced.