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.

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.

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. .

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.

Implantação e otimização da PCR em tempo real para o diagnóstico da influenza A (H1N1) pandêmica no Instituto Adolfo Lutz e perspectivas para 2010 / Implementation and optimization of real time PCR for diagnosis of pandemic influenza A (H1N1) at Instituto Adolfo Lutz and perspectives for 2010

Os autores apresentam sua experiência no diagnóstico laboratorial de InfluenzaA (H1N1) em 37.240 amostras clínicas obtidas de pacientes com suspeita de gripe, encaminhadas ao Instituto Adolfo Lutz para análise. Eles apresentam os algoritmos de testes moleculares empregados, comparam a eficiência dos mesmos quanto à sensibilidade, especificidade e custo e, finalmente sugerem um novo algoritmo para ser usado em caso de nova epidemia de Influenza A (H1N1) em 2010.

The authors present their experience with the molecular diagnosis of Influenza A (H1N1) with 37.240 clinicalsamples obtained from individuals suspected of flu, sent to Instituto Adolfo Lutz for analysis. They show the used algorithms, compare their efficiency in terms of sensitivity, specificity and cost, and suggest a new algorithm to be employed in case of an outbreak of Influenza A (H1N1) in 2010.

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.

Soroprevalência da hepatite B e avaliação da resposta imunológica à vacinação contra a hepatite B por via intramuscular e intradérmica em profissionais de um laboratório de saúde pública / Hepatitis B seroprevalence and evaluation of immune response to hepatitis B vaccination using intramuscular and intradermal routes in public health laboratory employees

OBJETIVOS: Determinar a prevalência dos marcadores da hepatite B (HBsAg e anti-HBs) e avaliar a resposta à vacinação contra hepatite B por via intradérmica (ID) em profissionais de saúde que não responderam à vacinação por via intramuscular (IM). MÉTODO: Todos os funcionários do Instituto Adolfo Lutz (IAL) foram convidados a participar do estudo. Amostras de soro foram colhidas no momento da administração da primeira dose de vacina (Engerix® B) e o HBsAg e o anti-HBs foram pesquisados, utilizando-se kits comerciais (Laboratórios Abbott®). Aos funcionários que não responderam à vacinação convencional (três doses por via IM) foram oferecidas doses de 5µg da mesma vacina por via ID. RESULTADOS: Foram envolvidos nesse estudo 404 funcionários do IAL, dos quais dois (0,5 por cento) eram HBsAg e 42 (10,5 por cento), anti-HBs reagentes. Dos 360 voluntários com sorologia negativa, 316 (87,8 por cento) receberam três doses de vacina (IM) e, desses, 259 colheram soros para avaliação pós-vacinal. Do total, 242 (93,4 por cento) apresentaram anticorpos acima de 10 UI/L após completarem o esquema inicial. Foram administradas duas doses de reforço, porém sete funcionários permaneceram sem resposta imunológica. A vacinação intradérmica foi realizada em cinco voluntários, e todos produziram anticorpos após a utilização dessa via de administração. CONCLUSÕES: A prevalência da hepatite B não foi maior nessa população do que na população geral. A vacinação por via intradérmica pode ser uma boa alternativa na imunização de pessoas que não respondem ao esquema convencional.

OBJECTIVES: To determine the prevalence of HBsAg and anti-HBs and to evaluate the response of intradermal hepatitis vaccination in healthcare workers non-responsive to previous repeated intramuscular vaccination. MATERIAL AND METHOD: All of the employees from Instituto Adolfo Lutz were invited to participate on this study. Serum samples were obtained and HBsAg and anti-HBs were detected using commercial kits (Abbott® Laboratories). Employees were submitted to the conventional three-dose vaccination by intramuscular route. To those employees who did not respond to intramuscular vaccination, 5 µg doses of Engerix® B were then administered by intradermal route up to nine doses. RESULTS: Four hundred and four healthcare workers were enrolled in this study. Initially, two (0.5 percent) and 42 (10.4 percent) were HBsAg and anti-HBs reagent, respectively. Among the 360 negative volunteers, 316 (87.8 percent) received three vaccine doses and in 259 of them, serum samples were collected to evaluate vaccine efficacy. Among them, 242 (93.4 percent) showed antibodies titer higher than 10 UI/l. Intradermal vaccination was carried out in five volunteers and all of them responded to this vaccine administration route. CONCLUSION: The prevalence of hepatitis B was not higher than in general population. Intradermal vaccine administration could be a good alternative in people that did not respond to previous intramuscular route.

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.

Duality of patterns in hepatitis a epidemiology: a study involving two socioeconomically distinct populations in Campinas, Sao Paulo State, Brazil

A avaliaçäo da prevalencia de anticorpos contra o virus da hepatite A em duas populaçöes com diferentes niveis sócio-economicos foi realizada pela analise de 101 e 82 amostras de soros provenientes de grupos de alto e baixo nivel sócio-economico, respectivamente, utilizando um teste imunoenzimatico comercial. A prevalência no grupo de baixo nivel sócio-economico foi 95,0 por cento enquanto que no grupo de alto nivel sócio-economico foi apenas 19,6 por cento (p < 0,001). Estes dados mostram uma dualidade no Brasil : a prevalência de anti-HAV em individuos de nivel sócio-economico baixo e similar aquela dos paises em desenvolvimento enquanto que nos individuos de alto nivel sócio-economico e compativel com o padrao de paises desenvolvidos. O controle desta infecçäo depende primariamente da melhoria das condiçöes sanitarias, mas especialmente em populaçöes de alto-nivel sócio-economico, o uso da vacinaçäo contra a hepatite A e altamente aconselhavel para evitar o aparecimento da doença em adultos