Characterization of primary direct-acting antiviral (DAA) drugs resistance mutations in NS5A/NS5B regions of hepatitis C virus with genotype 1a and 1b from patients with chronic hepatitis

ABSTRACT The Hepatitis C virus (HCV) infection is a public health problem. The high level of HCV replication and its lack of post-transcriptional correction mechanisms results in the emergence of viral variants and the difficulty in determining polymorphisms and variants that contain the substitutions associated with resistance towards new antivirals. The main focus of this study was to map the NS5A and NS5B polymorphisms and resistance mutations to new antiviral drugs in HCV strains genotype 1 from patients with chronic hepatitis C infection. Serum samples were collected from patients who underwent routine viral load tests at the Instituto Adolfo Lutz, Sao Paulo city, Brazil. A total of 698 and 853 samples were used for the characterization of NS5A and NS5B regions, respectively, which comprise the HCV genotypes 1a and 1b. The prevalence of resistance mutations found in the NS5A region was 6.4%, with Y93H, L31M, Q30R, and Y93N as the main resistance-associated substitutions (RAS). No NS5B-associated RAS was observed for any of the analyzed drugs. These findings support that the RAS test should be offered to individuals with poor response to double combination regimens prior to treatment initiation, thereby assisting strain vigilance and selection of effective treatment or retreatment options using DAA regimens.

Hepatitis B: Prevalence and occult infection in HIV-infected patients

Abstract INTRODUCTION: HBV and HIV have identical transmission routes. The aim of this study was to determine the prevalence of HBV in HIV patients and to detect the presence of occult HBV infection. METHODS: All samples were tested for serology markers and using qPCR. RESULTS: This study included 232 individuals, out of which 36.6% presented with HBV markers and 11.8% presented with HBsAg or HBV-DNA, including 3 patients that showed OBI. CONCLUSIONS: We observed a high prevalence of HBV among HIV patients. In addition, the results suggest that OBI can occur in patients with serological profiles that are indicative of past infection. Therefore, the application of molecular tests may enable the identification of infections that are not evident solely based on serology.

Surveillance of human retroviruses in blood samples from patients with hepatitis B and C in São Paulo, Brazil

Abstract INTRODUCTION Human retroviruses and the hepatitis B and C viruses (HBV and HCV, respectively) share routes of transmission; thus, coinfections occur and could alter subsequent disease outcomes. A preliminary study on human T-lymphotropic virus types 1 and 2 (HTLV-1/2) in serum samples from HBV- and HCV-infected individuals in São Paulo revealed 1.3% and 5.3% rates of coinfection, respectively. These percentages were of concern since they were detected in HTLV-endemic regions and in high-risk individuals in Brazil. The present study was conducted to extend and confirm these data. METHODS HTLV-1/2 and human immunodeficiency virus (HIV) infection status were identified in 1,984 sera for HBV and HCV viral load quantification - 1,290 samples from HBV-infected individuals (53.3% men, mean age: 47.1 years) and 694 samples from HCV-infected individuals (56.3% men, mean age: 50.1 years). HTLV-1/2 antibodies were detected by enzyme immunoassay, followed by western blotting and line immunoassay; HIV infection was detected by enzyme immunoassay. RESULTS HTLV-1/-2 infection was detected in 1.9% HBV-infected individuals (0.7% HTLV-1 and 1.2% HTLV-2) and in 4.0% (2.4% HTLV-1 and 1.6% HTLV-2) HCV-infected individuals; HIV infection was detected in 9.2% and 14.5%, respectively. Strong associations with HTLV and HIV, male sex, and older age were found in HBV/HTLV and HCV/HTLV-coinfected individuals (p<0.05). CONCLUSIONS HTLV-1 and HTLV-2 were confirmed to be prevalent in individuals with HBV and HCV in São Paulo; coinfected individuals deserve further clinical and laboratory investigation.

Fluctuations in serological hepatitis C virus levels in HIV patients

Abstract INTRODUCTION: Hepatitis C virus (HCV) and human immunodeficiency virus (HIV) have identical transmission routes, explaining the high prevalence of coinfections. The main aim of this study was to detect fluctuations in serological HCV levels in HIV patients. METHODS: We analyzed samples of 147 patients who attended an outpatient service that supports HIV/AIDS patients in São Paulo city. We also recruited 22 HCV-monoinfected patients who attended the Instituto Adolfo Lutz Laboratory in São Paulo city, to compare the test results. Serological testing of the blood samples was performed for the detection of HCV antibodies. The samples were then analyzed using real-time PCR for RNA viral quantification and sequencing. RESULTS We found that 13.6% of the study population was coinfected with HIV and HCV. In 20% of coinfected patients, fluctuations in serology results were detected in samples collected during the follow-up. No changes in anti-HCV serological markers were observed in HCV-monoinfected patients. An HCV viral load was detected in 9,5% of the samples collected from HIV patients. CONCLUSIONS: Our findings provide important clinical data to public health professionals and highlight the importance of periodic monitoring of HCV/HIV coinfected patients.

Hepatitis C viral load in HCV-monoinfected and HCV/HIV-1-, HCV/HTLV-1/-2-, and HCV/HIV/HTLV-1/-2-co-infected patients from São Paulo, Brazil

Co-infections of hepatitis C virus (HCV) and either human immunodeficiency virus type1 (HIV-1), human T-cell lymphotropic virus type 1 (HTLV-1) or type 2 (HTLV-2) have beendescribed as having an impact on HCV viremia and subsequent disease progression. HCVload in serum samples from 622 patients (343 males, 279 females; median age 50.8 years)from São Paulo/southeast Brazil was analyzed using the Abbott Real Time HCV assay(Abbott Molecular Inc., IL, USA). Samples were obtained from HCV-monoinfected (n = 548),HCV/HIV-1- (n = 41), HCV/HTLV-1- (n = 16), HCV/HTLV-2- (n = 8), HCV/HIV/HTLV-1- (n = 4), andHCV/HIV/HTLV-2-co-infected (n = 5) patients, and results were compared among the groupsand according to sex. The median HCV load in HCV-monoinfected patients was 5.23 log10IU/mL and 0.31 log10higher in men than in women. Increases in viral load of 0.51 log10, 0.54log10, and 1.43 log10IU/mL were detected in HCV/HIV-1-, HCV/HTLV-1- and HCV/HIV/HTLV-1-co-infected individuals, respectively, compared with HCV-monoinfected counterparts. Incontrast, compared to HCV/HIV co-infected patients, HCV/HTLV-2-co-infected patients hadan HCV load of 5.0 log10IU/mL, whereas HCV/HIV/HTLV-2-co-infected patients had a medianload 0.37 log10IU/mL lower. Significant differences in HCV loads were detected, with malesand HCV/HIV-1- and HCV/HIV/HTLV-1-co-infected patients presenting the highest values.Conversely, females and HCV/HTLV-2-co-infected patients exhibited lower HCV loads. Over-all, HCV viremia is increased in HIV and/or HTLV-1-co-infection and decreased in HTLV-2co-infection. (AU)

Hepatitis C viral load in HCV-monoinfected and HCV/HIV-1-, HCV/HTLV-1/-2-, and HCV/HIV/HTLV-1/-2-co-infected patients from São Paulo, Brazil

ABSTRACT Co-infections of hepatitis C virus (HCV) and either human immunodeficiency virus type 1 (HIV-1), human T-cell lymphotropic virus type 1 (HTLV-1) or type 2 (HTLV-2) have been described as having an impact on HCV viremia and subsequent disease progression. HCV load in serum samples from 622 patients (343 males, 279 females; median age 50.8 years) from São Paulo/southeast Brazil was analyzed using the Abbott Real Time HCV assay (Abbott Molecular Inc., IL, USA). Samples were obtained from HCV-monoinfected (n = 548), HCV/HIV-1- (n = 41), HCV/HTLV-1- (n = 16), HCV/HTLV-2- (n = 8), HCV/HIV/HTLV-1- (n = 4), and HCV/HIV/HTLV-2-co-infected (n = 5) patients, and results were compared among the groups and according to sex. The median HCV load in HCV-monoinfected patients was 5.23 log10 IU/mL and 0.31 log10 higher in men than in women. Increases in viral load of 0.51 log10, 0.54 log10, and 1.43 log10 IU/mL were detected in HCV/HIV-1-, HCV/HTLV-1- and HCV/HIV/HTLV-1-co-infected individuals, respectively, compared with HCV-monoinfected counterparts. In contrast, compared to HCV/HIV co-infected patients, HCV/HTLV-2-co-infected patients had an HCV load of 5.0 log10 IU/mL, whereas HCV/HIV/HTLV-2-co-infected patients had a median load 0.37 log10 IU/mL lower. Significant differences in HCV loads were detected, with males and HCV/HIV-1- and HCV/HIV/HTLV-1-co-infected patients presenting the highest values. Conversely, females and HCV/HTLV-2-co-infected patients exhibited lower HCV loads. Overall, HCV viremia is increased in HIV and/or HTLV-1-co-infection and decreased in HTLV-2 co-infection.

Making the invisible visible: searching for human T-cell lymphotropic virus types 1 and 2 (HTLV-1 and HTLV-2) in Brazilian patients with viral hepatitis B and C

With this study, the authors hope to alert clinicians regarding the presence of human T-cell lymphotropic virus type 1 and 2 (HTLV-1/-2) infections in patients with viral hepatitis B and C in Brazil. HTLV-1/-2 were detected in 1.3% of hepatitis B virus (HBV)- and 5.3% of hepatitis C virus (HCV)-infected blood samples sent for laboratory viral load measurements. A partial association of human immunodeficiency virus (HIV)-1 and HTLV-1/-2 infection was detected in patients with HCV (HIV+, 27.3%), whereas this association was almost 100% in HBV-infected patients (HIV+, all except one). The high prevalence of HTLV-1/-2 infection among patients with hepatitis C was of concern, as HTLV-1/-2 could change the natural course of subsequent liver disease. The authors suggest including HTLV-1/-2 serology in the battery of tests used when following patients with viral hepatitis in Brazil, regardless of the HIV status.

An in-house real-time polymerase chain reaction: standardisation and comparison with the Cobas Amplicor HBV monitor and Cobas AmpliPrep/Cobas TaqMan HBV tests for the quantification of hepatitis B virus DNA

This study aimed to standardise an in-house real-time polymerase chain reaction (rtPCR) to allow quantification of hepatitis B virus (HBV) DNA in serum or plasma samples, and to compare this method with two commercial assays, the Cobas Amplicor HBV monitor and the Cobas AmpliPrep/Cobas TaqMan HBV test. Samples from 397 patients from the state of São Paulo were analysed by all three methods. Fifty-two samples were from patients who were human immunodeficiency virus and hepatitis C virus positive, but HBV negative. Genotypes were characterised, and the viral load was measure in each sample. The in-house rtPCR showed an excellent success rate compared with commercial tests; inter-assay and intra-assay coefficients correlated with commercial tests (r = 0.96 and r = 0.913, p < 0.001) and the in-house test showed no genotype-dependent differences in detection and quantification rates. The in-house assay tested in this study could be used for screening and quantifying HBV DNA in order to monitor patients during therapy.

Identification of hepatitis B virus genotypes in the state of São Paulo / Identificação dos genótipos do vírus da hepatite B no Estado de São Paulo

Objective: the aim of this study was to identify HBV genotypes in serum samples from patients from the state of São Paulo, received by the viral hepatitis laboratory, at the Virology Centre of Instituto Adolfo Lutz, from various municipalities. Methods: a total of 94 serum samples were randomly analyzed. Genotyping was performed using nested PCR for amplification of S and Pol regions from viral genome. Genotypes were identified comparing the sequences obtained with the sequences deposited in GenBank. Results: we were able to determine the genotype of 91 (97%) samples, as follows: genotype A (55.3%), D (32%), F (5.3%), C (3.2%) and G (1%). There are few data on the epidemiology of genotype G. This genotype has been detected in restricted areas around the world. Frequently, the genotype G infection occurs in HIV-positive male patients. In our case, the sample identified as G was also positive for HIV but in a female patient, which is an uncommon finding in the scientific literature. Conclusion: in this work, we identified the most frequent genotypes in São Paulo as well as the genotype G, rare among the genotypes found in our environment. .

Objetivo: o objetivo deste estudo foi identificar os genótipos do HBV nas amostras de soros recebidas pelo Laboratório de Hepatites do Centro de Virologia do Instituto Adolfo Lutz. Métodos: foram analisadas aleatoriamente 94 amostras de soropositivas, provenientes de diversos municípios do Estado de São Paulo. Para determinação dos genótipos, foi realizada Nested-PCR das regiões S e Pol do HBV. Os genótipos foram identificados comparando os resultados amplificados com as sequências depositadas no GenBank. Resultados: foi possível determinar o genótipo de 91 (97%) amostras do total analisado e os genótipos identificados foram: genótipos A (55,3%), D (32%), F (5,3%), C (3,2%) e G (1%). Há poucos dados a respeito da epidemiologia do genótipo G. Esse genótipo tem sido detectado em áreas restritas do mundo. Geralmente, a infecção pelo genótipo G ocorre em indivíduos HIV positivos do sexo masculino. Neste trabalho, a amostra identificada como G foi também positiva para HIV e era de uma paciente do sexo feminino, dado raro na literatura científica. Conclusão: neste trabalho, foram identificados os genótipos mais frequentes, assim como uma cepa do genótipo G, rara entre os encontrados em nosso meio. .

Epidemiological, serological and molecular aspects of hepatitis B and C in children and teenagers of municipal daycare facilities schools and schools in the city of Santos / Aspectos epidemiológicos, sorológicos e moleculares das hepatites B e C em crianças e adolescentes de creches e escolas municipais na cidade de Santos

Objective: To estimate the prevalence of the serological markers anti-HBc, HBsAg and anti-HBs of hepatitis B and anti-HCV of hepatitis C among children and teenagers enrolled at daycare facilities, kindergartens and municipal elementary education network in the city of Santos, São Paulo, Brazil. Methods: A cross-sectional study was carried out from June 28 to December 14, 2007, in which 4,680 finger-prick blood samples were collected from children and teenagers. A survey questionnaire was applied to their family members. The sample was dimensioned using the software Epi Info version 6 with expected frequency of 1%, acceptable error of 0.5% and confidence interval of 95%. The serological tests were performed using the ELISA technique. The molecular analysis was performed using the technique of polymerase chain reaction in House. Results: Age of the studied population ranged from 7 months to 18 years and 1 month. The general prevalence of anti-HBc reagent was 0.1%, HBsAg was 0.02% and anti-HCV was 0.02%. Conclusions: In children, the general prevalence of serological markers for hepatitis B and C in the city of Santos was low when compared with literature data. .

Objetivo: Estimar a prevalência de marcadores sorológicos anti-HBc, AgHBs e anti-HBs da hepatites B e anti-HCV da hepatite C em crianças e adolescentes matriculados em creches e escolas de ensino infantil e fundamental da rede municipal na cidade de Santos, São Paulo. Métodos: Estudo transversal realizado no período de 28 de junho a 14 de dezembro de 2007, no qual foram coletadas 4.680 amostras de sangue colhidas através de punção capilar. Foi aplicado um questionário nos familiares das crianças e adolescentes. Para o cálculo da amostra, foi utilizado o programa Epi Info versão 6 com frequência esperada de 1%, erro aceitável de 0,5% e nível de confiança de 95%. Os exames sorológicos foram realizados utilizando a técnica de ELISA. O estudo molecular foi realizado pela técnica de reação em cadeia de polimerase in House. Resultados: A idade da população estudada variou de 7 meses a 18 anos e 1 mês. A prevalência geral do anti-HBc reagente foi de 0,1%, do AgHBs foi de 0,02% e do anti-HCV foi de 0,02%. Conclusão: A prevalência geral em crianças dos marcadores sorológicos para hepatites B e C na cidade de Santos foi baixa, quando comparada com os dados de literatura. .

Occult hepatitis B virus infection in hemodialysis patients in Recife, State of Pernambuco, Brazil / Infecção oculta para o vírus da hepatite B em pacientes hemodialisados de Recife, Estado de Pernambuco, Brasil

INTRODUCTION: Persistence of the hepatitis B virus (HBV) genome in individuals negative for the HBV surface antigen (HBsAg) reflects occult infection. The aim of this study was to identify occult HBV infection among hemodialysis patients at 5 clinics in Recife, State of Pernambuco, Brazil, between August 2006 and August 2007. METHODS: Serum samples underwent enzyme-linked immunosorbent assay to investigate total antibodies against HBcAg (anti-HBc), HBsAg, and antibodies against HBsAg (anti-HBs). Samples that were HBsAg-negative were tested for total anti-HBc, and those that were positive for total anti-HBc were tested for anti-HBs. HBV DNA was investigated with an in-house PCR technique to identify samples positive for total anti-HBc. Subsequently, the samples positive for HBV DNA were sequenced to identify the genotype and mutations. RESULTS: The study population (n = 752) had a mean age of 50 15.1 years and included both sexes. All samples analyzed were negative for HBsAg. The seroprevalence of total anti-HBc was 26.7% (201/752), while that of anti-HBs was 67.2% (135/201). Total anti-HBc alone was detected in 5.7% of the patients. Occult infection was found in 1.5%, comprising genotypes A (33.3%, 1/3) and D (66.7%, 2/3). No mutations were found. CONCLUSIONS: The study detected occult hepatitis B virus infection in hemodialysis patients. Molecular studies on HBV are of fundamental importance because they identify patients that had been considered virus-negative but who, in reality, host the virus and have the ability to transmit it to other patients and staff.

INTRODUÇÃO: A persistência do genoma do vírus da hepatite B (HBV) em indivíduos com o antígeno de superfície para o HBV (HBsAg) negativo, caracteriza-se como infecção oculta. O objetivo desse trabalho foi identificar infecção oculta pelo HBV em hemodialisados provenientes de cinco clínicas de Recife, Estado de Pernambuco, Brasil, de agosto de 2006 a agosto de 2007. MÉTODOS: Os soros foram submetidos à pesquisa do HBsAg, anticorpos totais para o AgHBc (anti-HBc total) e para os anticorpos contra o antígeno de superfície do HBV (anti-HBs) pelo ensaio imunoenzimático (ELISA). As amostras negativas para o HBsAg foram testadas para o anti-HBc total e os soros reativos, foram testados para o anti-HBs. O DNA-HBV foi analisado pela reação em cadeia da polimerase (PCR) in house nas amostras anti-HBc total reagentes. As amostras DNA-HBV positivas foram sequenciadas para a identificação do genótipo e pesquisa de mutação. RESULTADOS: A população estudada foi de 752 hemodialisados, com média de idade de 50±15,1 e ambos os sexos. Todos os soros analisados foram HBsAg negativos. A soroprevalência para o anti-HBc total e anti-HBs foi de 26,7% (201/752) e 67,2% (135/201), respectivamente. O anti-HBc total foi encontrado em 5,7% dos pacientes. A infecção oculta foi identificada em 1,5% (3/201) das amostras, sendo 33,3% com o genótipo A e 66,7% com o D. Nenhuma mutação foi observada. CONCLUSÕES: O estudo encontrou infecção oculta na amostra analisada. A infecção oculta encontrada mostra a importância de estudos moleculares, pois pacientes HBsAg negativos podem albergar o vírus disseminando-o para os demais pacientes.

Hepatitis B virus infection in children, adolescents, and their relatives: genotype distribution and precore and core gene mutations / Infecção pelo vírus da hepatite B em crianças, adolescentes e seus familiares: distribuição dos genótipos e mutações no gene pré-core e core

INTRODUCTION:The objectives of this study were evaluate hepatitis B virus (HBV) serological markers in children and adolescents followed up at the Child Institute of the Hospital das Clínicas, Faculdade de Medicina de São Paulo, Universidade de São Paulo; identify chronic HBV carriers and susceptible individuals in the intrafamilial environment; characterize HBV genotypes; and identify mutations in the patients and household contacts. METHODS: Ninety-five hepatitis B surface antigen-positive children aged <19 years and 118 household contacts were enrolled in this study. Commercial kits were used for the detection of serological markers, and PCR was used for genotyping. RESULTS: Hepatitis B e antigen (HBeAg) was detected in 66.3% (63/95) of cases. Three of the 30 HBeAg-negative and anti-HBeAg-positive patients presented with precore mutations and 11 presented with mutations in the basal core promoter (BCP). Genotype A was identified in 39 (43.8%) patients, genotype D in 45 (50.6%), and genotype C in 5 (5.6%). Of the 118 relatives, 40 were chronic HBV carriers, 52 presented with the anti-HBc marker, 19 were vaccinated, and 7 were susceptible. Among the relatives, genotypes A, D, and C were the most frequent. One parent presented with a precore mutation and 4 presented with BCP mutations. CONCLUSIONS: Genotypes A and D were the most frequent among children, adolescents, and their relatives. The high prevalence of HBV in the families showed the possibility of its intrafamilial transmission.

INTRODUÇÃO: Os objetivos deste estudo foram: avaliar os marcadores sorológicos nas crianças e adolescentes acompanhadas no Instituto da Criança do Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo (HC-FMUSP); identificar portadores crônicos do VHB e indivíduos suscetíveis no ambiente intrafamiliar; caracterizar o genótipo do VHB; observar a presença de cepas mutantes entre os pacientes e familiares estudados. MÉTODOS: Noventa e cinco crianças e adolescentes positivas para o antígeno de superfície do vírus da Hepatite B (AgHBs), menores de 19 anos, e 118 familiares foram envolvidos neste estudo. Foram utilizados kits comerciais para a pesquisa dos marcadores sorológicos e a PCR foi utilizada para genotipagem. RESULTADOS: O antígeno e do vírus da hepatite B (AgHBe) foi detectado em 66,3% (63/95) dos casos. Três dos 30 pacientes AgHBe negativo e anti-HBe positivo apresentaram mutação na região pré-core e 11 na região BCP. Em 39 (43,8%) pacientes, foi identificado o genótipo A, 45 (50,6%)o genótipo D e cinco (5,6%) o genótipo C. Dos 118 familiares estudados, 40 eram portadores crônicos do VHB, 52 tinham marcador sorológico de contato prévio e sete eram suscetíveis. Dentre os familiares AgHBs positivos, os genótipos A, D e C foram os mais frequentes. Um familiar apresentou mutação na região pré-core e quatro apresentaram mutação na região do BCP. CONCLUSÕES: Os genótipos A e D foram os mais frequentes dentre as crianças adolescentes e seus familiares. Alta frequência do VHB nos familiares mostrou a possibilidade de transmissão intrafamiliar.

Epidemiological and serological aspects of hepatitis A among children and teenagers in the city of Santos: a cross-sectional study / Aspectos epidemiológicos e sorológicos da hepatite A em crianças e adolescentes na cidade de Santos: estudo transversal

CONTEXT AND OBJECTIVE: Viral hepatitis A is still a concern at public health level in Brazil and around the world, due both to the number of affected subjects and the possibility of complications in the acute forms. The Brazilian Ministry of Health estimates that at least 70% of this country's population has already had contact with the hepatitis A virus (HAV). The aim here was to discover the prevalence of serological markers for the hepatitis A virus among children and teenagers at daycare facilities, kindergartens and elementary schools in the city of Santos. DESIGN AND SETTING: Cross-sectional study in kindergartens and elementary schools within the municipal education network in several regions of the city of Santos. METHOD: Students' family members were surveyed using a questionnaire and 4,680 finger-prick blood samples were taken and assayed by means of the ELISA technique. RESULTS: The general prevalence of anti-HAV IgG was 9.72% and, of these cases, 74.6% were reactive to anti-HAV IgM. There was higher prevalence of anti-HAV IgG among older children, females, children who played in streams, those whose homes were not connected to the sewage system, those whose parents had low education levels, those with low household income and those who did not live along the seashore. The prevalence of anti-HAV IgM peaked in the early years and subsequently fell, and it was lower on the hills and in the Northwestern Zone. CONCLUSION: The general prevalence of serological markers for hepatitis A was low in Santos.

CONTEXTO E OBJETIVO: A hepatite viral A continua sendo uma preocupação em nível de saúde pública no Brasil e no mundo, tanto pelo número de indivíduos atingidos, como pela possibilidade de complicação das formas agudas. O Ministério da Saúde estima que pelo menos 70% da população do Brasil já tiveram contato com o vírus da hepatite A. O objetivo foi conhecer a prevalência de marcadores sorológicos do vírus da hepatite A em crianças e adolescentes de creches e escolas de ensino infantil e fundamental na cidade de Santos. TIPO DE ESTUDO E LOCAL: Estudo transversal em pré-escolas e de ensino fundamental da rede municipal em diversas regiões da cidade de Santos. MÉTODO: Foi aplicado um questionário aos familiares dos estudantes e coletadas 4.680 amostras de sangue através de punção capilar para realização da sorologia pela técnica ELISA. RESULTADOS: A prevalência geral do anti-HVA IgG foi de 9,72% e, desses, 74,6% foram anti-HVA IgM reagentes. A prevalência de anti-HVA IgG foi maior entre as crianças mais velhas, meninas, aquelas que brincavam em córregos, sem rede de coleta de esgoto em sua moradia, de pais com baixa instrução, de baixa renda familiar e aquelas que não eram moradoras da orla. A prevalência de anti-HVA IgM teve pico nos primeiros anos e posterior queda e, no morro e Zona Noroeste, foi mais baixa. CONCLUSÃO: A prevalência geral dos marcadores sorológicos para hepatite A foi baixa em Santos.

Hepatitis B genotype G and high frequency of lamivudine-resistance mutations among human immunodeficiencyvirus/hepatitis B virus co-infected patients in Brazil

In this study, we evaluated the hepatitis B virus (HBV) genotype distribution and HBV genomic mutations among a group of human immunodeficiency virus-HBV co-infected patients from an AIDS outpatient clinic in São Paulo. HBV serological markers were detected by commercially available enzyme immunoassay kits. HBV DNA was detected using in-house nested polymerase chain reaction and quantified by Cobas Amplicor. HBV genotypes and mutations in the basal core promoter (BCP)/pre-core/core regions and surface/polymerase genes were determined by sequencing. Among the 59 patients included in this study, 55 reported prior use of lamivudine (LAM) or tenofovir. HBV DNA was detected in 16/22 patients, with a genotype distribution of A (n = 12,75 percent), G (n = 2,13 percent), D (n = 1,6 percent) and F (n = 1,6 percent). The sequence data of the two patients infected with genotype G strongly suggested co-infection with genotype A. In 10 patients with viremia, LAM-resistance mutations in the polymerase gene (rtL180M + rtM204V and rtV173L + rtL180M + rtM204V) were found, accompanied by changes in the envelope gene (sI195M, sW196L and sI195M/sE164D). Mutations in the BCP and pre-core regions were identified in four patients. In conclusion, genotype G, which is rarely seen in Brazil, was observed in the group of patients included in our study. A high prevalence of mutations associated with LAM-resistance and mutations associated with anti-HBs resistance were also found among these patients.

HBV markers in haemodialysis Brazilian patients: a prospective 12-month follow-up

The aim of this study was to determine the prevalence and the incidence of hepatitis B virus (HBV) among haemodialysis (HD) subjects and to evaluate whether testing for serological markers at the time of admission is suitable for HBV screening in this population. One hundred twenty-three patients belonging to two HD centres from São Paulo, Brazil, were tested prospectively. HBV DNA was detected by polymerase chain reaction (PCR) in each of the prospective subjects (n = 123) during one year. Additionally, all samples (n = 1,476) were analysed for HBV serological markers. The prevalence of hepatitis B core antibody (anti-HBc), hepatitis B surface antigen (HBsAg) and HBV DNA were 34.1 percent, 15.4 percent and 8.1 percent, respectively, while the incidence was null. Fluctuation in HBV serology was observed in one patient. Only 37.8 percent (17/45) of cases responded to the HBV vaccine. Our results suggest that employing more than one HBV marker and repeated follow-up evaluations may improve HBV screening in HD units.

Characterization of a Hepatitis B virus strain in southwestern Paraná, Brazil, presenting mutations previously associated with anti-HBs Resistance / Caracterização de uma cepa de hepatite por vírus B no sudoeste do Paraná, Brasil, apresentando mutações previamente associadas à resistência anti-HBs

The present study investigated if hepatitis B virus (HBV) mutants circulate in the southwestern region of the State of Paraná, Brazil, by analyzing samples from children who received immunoprophylaxis but were born to HBV carrier mothers. Samples from 25 children were screened for HBV serum markers and for HBV DNA by PCR. Only one sample was positive for HBsAg, anti-HBs and HBV DNA, although the child had been vaccinated. Analysis of the S gene sequence of this sample showed the presence of a proline at position 105, a serine at position 114, three threonines at positions 115, 116 and 140, and a glutamine at position 129. The presence of these amino acids, except for serine at position 114, has been related to monoclonal or polyclonal therapy with anti-HBs after liver transplantation, whereas the presence of threonine at position 116 has been described in immunized children from Singapore. This finding demonstrates the possible circulation of HBV strains resistant to hepatitis B immunoprophylaxis in southwestern Paraná, Brazil. The genotype of the sample was identified as genotype D, which is frequently found in the region studied. Since 36 percent of the children had received incomplete or no immunoprophylaxis, more extensive follow-up of children born to HBsAg-positive mothers is needed.

O presente estudo investigou se mutantes do vírus da hepatite B (HBV) circulam na região Sudoeste do Estado do Paraná, Brasil, analisando amostras de crianças que receberam a imunoprofilaxia por terem nascido de mães portadoras do HBV. Amostras de 25 crianças foram analisadas para os marcadores sorológicos do HBV e para o DNA-HBV por PCR. Somente uma amostra foi positiva para AgHBs, anti-HBs e DNA-HBV, apesar da criança ter sido vacinada. Análises da seqüência do gene S desta amostra mostrou a presença de uma prolina na posição 105, uma serina na posição 114, três treoninas nas posições 115, 116 e 140, e uma glutamina na posição 129. A presença destes aminoácidos, exceto para Serina na posição 114, tem sido relacionada a terapia monoclonal ou policlonal com anti-HBs após transplante de fígado, enquanto a presença da treonina na posição 116 tem sido descrita em crianças imunizadas de Singapura. Este achado demonstra a possível circulação de cepas do HBV resistentes a imunoprofilaxia para hepatite B no Sudoeste do Paraná, Brasil. O genótipo da amostra foi identificado como genótipo D, o qual é frequentemente encontrado na região estudada. Desde que 36 por cento das crianças tinham recebido incompleta ou nenhuma imunoprofilaxia, um seguimento mais intensivo das crianças nascidas de mães AgHBs positivo é necessário.

Hepatitis C and hemodialysis: a review

Hepatitis C is a serious public health problem throughout the world; chronic renal patients are highly exposed to this infection. This could be due to a failure to identify carriers of this disease or because of a lack of truly effective biosafety measures implemented in the dialysis units. Molecular biology techniques have allowed for the understanding of this virus in detail, including its replication mechanisms. Epidemiological studies have been made throughout the world, with the goal of determining the dissemination dynamics of this agent, in addition to examining the predominance of the different genotypes, and the possible mutants that are involved. Many questions must still be answered concerning infection by Hepatitis C virus (HCV); this is especially important for immunosuppressed patients.

Prevalence and risk factors of hepatitis C virus infection in hemodialysis patients from one center in Recife, Brazil

A hemodialysis population from a dialysis unit in the city of Recife, Northeastern Brazil, was screened to assess the prevalence of hepatitis C virus (HCV) infection and to investigate the associated risk factors. Hemodialysis patients (n = 250) were interviewed and serum samples tested for anti-HCV antibodies by enzyme-linked immunosorbent assay (ELISA). All samples were also tested for HCV RNA by reverse transcriptase nested polymerase chain reaction (RT-nested-PCR). Out of 250 patients, 21 (8.4 percent) were found to be seropositive by ELISA, and 19 (7.6 percent) patients were HCV RNA positive. HCV viraemia was present in 90.5 percent of the anti-HCV positive patients. The predominant genotype was HCV 1a (8/19), followed by 3a (7/19), and 1b (4/19). None of the anti-HCV negative patients were shown to be viraemic by the PCR. Univariate analysis of risk factors showed that time spent on hemodialysis, the number of blood transfusions and a blood transfusion before November 1993 were associated with HCV positivity. However, multivariate analysis revealed that blood transfusions before November 1993 were significantly associated with HCV infection in this population. Low prevalence levels were encountered in this center, however prospective studies are necessary to confirm these findings.

Detection of hepatitis A antibodies by ELISA using saliva as clinical samples

The possibility of detecting acute infection and immunity using body fluids that are easier to collect than blood, mainly in children, would facilitate the investigation and follow-up of outbreaks of hepatitis A (HAV). Our study was carried out to evaluate the detection of anti-HAV IgM, IgA and total antibodies in saliva using serum samples as reference. Forty three paired serum and saliva samples were analyzed. From this total, 24 samples were obtained from children and 1 from one adult during the course of acute hepatitis A; an additional 18 samples were obtained from health professionals from Adolfo Lutz Institute. The sensitivity to detect anti-HAV IgM was 100 per cent (95 per cent CI: 79.1 to 100.0 per cent), employing saliva as clinical samples. In detecting anti-HAV IgA, the sensitivity was 80.8 per cent (95 per cent CI: 60.0 to 92.7 per cent) and for the total antibodies was 82.1 per cent (95 per cent CI: 62.4 to 93.2 per cent). The specificity was 100 per cent for each. The rate of agreement was high comparing the results of serum and saliva samples for detecting HAV antibodies. We conclude that saliva is an acceptable alternative specimen for diagnosing acute hepatitis A infection, and for screening individuals to receive hepatitis A vaccine or immunoglobulin.