Hepatitis B in the Northwestern region of Sao Paulo State: genotypes and resistance mutations.

In Brazil, few studies on the molecular aspects of hepatitis B virus (HBV) infection have been conducted in the interior regions of Sao Paulo State. This study aimed to identify HBV genotypes and evaluate strains with resistance mutations for nucleoside analogues in the Administrative Region (AR) of the municipality of Sao Jose do Rio Preto. We performed nested PCRs of 127 samples from the Health Care Services of the AR to amplify, sequence and analyze fragments of the HBV DNA, in order to identify genotypes and resistance mutations. The HBV S/Pol regions of 126 samples were successfully amplified and sequenced. Five different genotypes were found, and the main ones were A, D and F; a greater number of samples contained the subgenotypes A1 (n = 51; 40.5%), D3 (n = 36; 28.6%), A2 (n = 14; 11.1%) and F2a (n = 9; 7.1%). Resistance mutations (rtM204V/I/S) associated or not with compensatory mutations (rtL180M, rtV173L) were identified in 13.9% (5/36) of patients undergoing viral treatment and 1.1% (1/90) of naïve patients. The diversity of genotypes/subgenotypes found is probably due to the intense migration occurring in the region. These data can complement epidemiological and clinical surveillance, and can be used for a more effective management of chronic HBV patients.

Occurrence of hepatitis B in pregnant women and follow-up of exposed children in the State of São Paulo, Brazil, in 2012. / Ocorrência de hepatite B em gestantes e seguimento de crianças expostas no estado de São Paulo, em 2012.

OBJECTIVE: to describe the occurrence of hepatitis B among pregnant women, immunoprophylaxis and vertical and perinatal transmission in children exposed to the virus in the São Paulo state primary care network, Brazil. METHODS: this was a cross-sectional study using prenatal records of pregnant women attending health services between January and June 2012 and a cohort of newborns; the frequencies of the results were described and the estimated occurrence of hepatitis B was calculated. RESULTS: 6,233 pregnant women were included, of whom 53.1% were between 20-29 years old, 58.7% had 8-11 years of schooling, 53.3% were white, and 73.9% lived with a partner; occurrence of hepatitis B was 0.13% (95%CI 0.04; 0.21); of the eight children of mothers with chronic hepatitis B, six had a complete vaccination schedule, and there was no vertical or perinatal transmission. CONCLUSION: there was low occurrence of hepatitis B in pregnant women and absence of vertical or perinatal transmission.

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

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.

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.

Ocorrência de hepatite B em gestantes e seguimento de crianças expostas no estado de São Paulo, em 2012 / Ocurrencia de hepatitis B en mujeres embarazadas y seguimiento de niños expuestos en el estado de São Paulo, Brasil, en 2012 / Occurrence of hepatitis B in pregnant women and follow-up of exposed children in the State of São Paulo, Brazil, in 2012

Resumo Objetivo: descrever a ocorrência da hepatite B entre gestantes, a realização de imunoprofilaxia e a transmissão vertical e perinatal nas crianças expostas ao vírus na rede de Atenção Primária à Saúde do estado de São Paulo, Brasil. Métodos: estudo transversal sobre registros de pré-natal de gestantes atendidas de janeiro a junho de 2012 e coorte dos recém-nascidos; foram descritas as frequências dos resultados e calculada a estimativa da ocorrência da hepatite B. Resultados: foram incluídas 6.233 gestantes, das quais 53,1% com 20 a 29 anos de idade, 58,7% com 8 a 11 anos de estudo, 53,3% brancas e 73,9% com companheiro; a ocorrência de hepatite B foi de 0,13% (IC95%: 0,04 a 0,21%); das oito crianças de mães com hepatite B crônica, seis tiveram esquema vacinal completo e não houve transmissão vertical ou perinatal. Conclusão: observou-se baixa ocorrência de hepatite B em gestantes e ausência de transmissão vertical ou perinatal.

Resumen Objetivo: describir el aparecimiento de hepatitis B en mujeres embarazadas, la inmunoprofilaxis y la transmisión vertical y perinatal en niños expuestos al virus en la red de atención primaria en el estado de São Paulo, Brasil. Métodos: estudio transversal con registros prenatales de mujeres embarazadas atendidas de enero a junio de 2012 y cohorte de recién nacidos; se describieron las frecuencias y se calculó el surgimiento de la hepatitis B. Resultados: se incluyeron 6.233 gestantes, de las cuales 53,1% con 20 a 29 años de edad, 58,7% con 8 a 11 años de estudios, 53,3% blancas y 73,9% viviendo en pareja; la ocurrencia de hepatitis B fue del 0,13% (IC95%: 0,04 to 0,21%); entre ocho hijos de madres con hepatitis B crónica, seis tenía un calendario de vacunación completo y no había transmisión vertical o perinatal. Conclusión: hubo baja ocurrencia de hepatitis B en mujeres embarazadas y ausencia de transmisión vertical o perinatal.

Abstract Objective: to describe the occurrence of hepatitis B among pregnant women, immunoprophylaxis and vertical and perinatal transmission in children exposed to the virus in the São Paulo state primary care network, Brazil. Methods: this was a cross-sectional study using prenatal records of pregnant women attending health services between January and June 2012 and a cohort of newborns; the frequencies of the results were described and the estimated occurrence of hepatitis B was calculated. Results: 6,233 pregnant women were included, of whom 53.1% were between 20-29 years old, 58.7% had 8-11 years of schooling, 53.3% were white, and 73.9% lived with a partner; occurrence of hepatitis B was 0.13% (95%CI 0.04; 0.21); of the eight children of mothers with chronic hepatitis B, six had a complete vaccination schedule, and there was no vertical or perinatal transmission. Conclusion: there was low occurrence of hepatitis B in pregnant women and absence of vertical or perinatal transmission.

Fluctuations in serological hepatitis C virus levels in HIV patients.

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.

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.

Diagnóstico Molecular da Hepatite C dos pacientes atendidos pelo Instituto Adolfo Lutz Central, entre março de 2012 e março de 2013 / Molecular Diagnosis of Hepatitis C patients attended by Institute Adolfo Lutz Central, from March 2012 to March 2013

O estudo avaliou a rotina do laboratório de Hepatites Virais do Laboratório Central do Instituto Adolfo Lutz - IAL Central, coordenador da Rede Estadual de Diagnóstico Molecular de São Paulo, no período de março de 2012 a março de 2013. O IAL Central atende a demanda das Regiões do ABCD, Vale do Paraíba, Vale do Ribeira, Franco da Rocha e na capital os atendimentos do Hospital São Paulo (Unifesp). Os exames realizados foram a quantificação da carga viral do VHC (HCVQT) por Real Time PCR Abbott® e a genotipagem (HCVG) utilizando o kit Versant® HCV genotype Inno-LiPA Assay (Siemens). Foram recebidas e analisadas 7.377 amostras no período, sendo que 12,2% dos pedidos foram cancelados, principalmente por: cadastro indevido, qualidade da amostra, volume insuficiente de amostra para realizar o teste. Dos exames realizados, a proporção de amostras encaminhadas pelas regiões foram: ABCD e município de São Paulo (50,9%), Vale do Paraíba (26,3%), Campinas (8,6%), Botucatu (7,7%), Franco da Rocha (6,1%) e Vale do Ribeira (0,4%). Os pacientes apresentaram mediana de idade de 49 anos e 57% eram do sexo masculino. O genótipo mais frequente foi o um (1) com 71,3%. Já o teste HCVQT apresentou mediana de carga viral de 3,28 log. (1.933,5UI/mL). O tempo médio de liberação de exames foi de 16 dias para HCVG e 12 para HCVQT. Os resultados obtidos permitiram conhecer a epidemiologia molecular das regiões atendidas pelo IAL e reforçaram a importância da Rede no tratamento dos infectados. Além disto, as falhas observadas ressaltam o papel do IAL Central em orientar e coordenar os laboratórios participantes da Rede de Diagnóstico Molecular...

Hepatitis B virus infection in children, adolescents, and their relatives: genotype distribution and precore and core gene mutations.

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.

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.

Estudo da infecção pelo vírus da hepatite B em crianças e adolescentes e seus familiares: distribuição dos genótipos e pesquisa de mutações no gene pré-core e core / Study of hepatitis B virus infection in children, adolescent and their relatives: genotypes distribuition and precore and core gene mutation

A infecção pelo vírus da hepatite B (VHB) na infância representa um desafio, pois as crianças que adquirem o VHB, por transmissão perinatal, têm risco de 90% de desenvolver infecção crônica. Vários estudos têm sido realizados na tentativa de demonstrar a relação entre os genótipos e a evolução clínica desta infecção, porém estes são ainda escassos e pouco consistentes, principalmente no que diz respeito à hepatite B na infância, devido ao número pequeno de pacientes envolvidos. Este estudo teve por objetivos caracterizar a infecção pelo VHB em crianças e adolescentes acompanhadas no Instituto da Criança do HC/FMUSP; avaliar os marcadores sorológicos; caracterizar o genótipo do VHB e a presença de mutações da região Pré-core e do BCP; identificar portadores crônicos do VHB e indivíduos suscetíveis no ambiente intrafamiliar; caracterizar o genótipo do VHB e observar a presença de cepas mutantes entre os familiares. A população desse estudo foi constituída por 95 crianças e adolescentes, menores de 19 anos, que apresentaram AgHBs positivo por mais de seis meses e 118 familiares dos pacientes. Foram utilizados kits comerciais para a pesquisa dos marcadores sorológicos. A “nested” PCR para região S foi utilizada para genotipagem do VHB nas amostras com AgHBs. Nas amostras AgHBs/anti-HBe positivas e AgHBe negativas foram pesquisadas mutações empregando-se a “nested” PCR para região précore. Das 95 crianças e adolescentes, 53 eram do gênero masculino e 42 do feminino, com média de idade de 7,6 anos. Em relação à sorologia, 63 (66,3%) apresentaram AgHBe e 2,1% (2/95) não apresentaram anti-HBc total. Das 32 amostras AgHBe negativas, 30 (93,7%) apresentaram resultado positivo para o anti-HBe, dos quais três apresentaram mutação na região pré-core e 11 na região BCP. Das 89 amostras de pacientes em que foi possível amplificar a região S, 39 (43,8%) foram caracterizadas como genótipo A, cinco (5,6%) como C e 45 como D (50,6%). Dos 118 familiares estudados, 40 eram portad...

Estudo da infecção pelo vírus da hepatite B em crianças e adolescentes e seus familiares: distribuição dos genótipos e pesquisa de mutações no gene pré-core e core

A infecção pelo vírus da hepatite B (VHB) na infância representa um desafio, pois as crianças que adquirem o VHB, por transmissão perinatal, têm risco de 90% de desenvolver infecção crônica. Vários estudos têm sido realizados na tentativa de demonstrar a relação entre os genótipos e a evolução clínica desta infecção, porém estes são ainda escassos e pouco consistentes, principalmente no que diz respeito à hepatite B na infância, devido ao número pequeno de pacientes envolvidos. Este estudo teve por objetivos caracterizar a infecção pelo VHB em crianças e adolescentes acompanhadas no Instituto da Criança do HC/FMUSP; avaliar os marcadores sorológicos; caracterizar o genótipo do VHB e a presença de mutações da região Pré-core e do BCP; identificar portadores crônicos do VHB e indivíduos suscetíveis no ambiente intrafamiliar; caracterizar o genótipo do VHB e observar a presença de cepas mutantes entre os familiares. A população desse estudo foi constituída por 95 crianças e adolescentes, menores de 19 anos, que apresentaram AgHBs positivo por mais de seis meses e 118 familiares dos pacientes. Foram utilizados kits comerciais para a pesquisa dos marcadores sorológicos. A “nested” PCR para região S foi utilizada para genotipagem do VHB nas amostras com AgHBs. Nas amostras AgHBs/anti-HBe positivas e AgHBe negativas foram pesquisadas mutações empregando-se a “nested” PCR para região précore. Das 95 crianças e adolescentes, 53 eram do gênero masculino e 42 do feminino, com média de idade de 7,6 anos. Em relação à sorologia, 63 (66,3%) apresentaram AgHBe e 2,1% (2/95) não apresentaram anti-HBc total. Das 32 amostras AgHBe negativas, 30 (93,7%) apresentaram resultado positivo para o anti-HBe, dos quais três apresentaram mutação na região pré-core e 11 na região BCP. Das 89 amostras de pacientes em que foi possível amplificar a região S, 39 (43,8%) foram caracterizadas como genótipo A, cinco (5,6%) como ...

High prevalence of hepatitis B virus and hepatitis D virus in the western Brazilian Amazon.

Severe cases of hepatitis caused by hepatitis B virus (HBV) or hepatitis D virus (HDV) are often seen in the Brazilian Amazon, but there is a paucity of epidemiologic studies on viral hepatitis in this area. Thus, a cross-sectional study to investigate the prevalence of markers for HBV and HDV was performed. Serum samples were collected after participants completed an epidemiologic questionnaire. Markers for HBV and HDV were analyzed with an enzyme-linked immunosorbent assay. The HBV genotype was determined by sequencing of the gene for hepatitis B surface antigen (HBsAg). Of 2,656 samples, 89 (3.3%) were positive for HBsAg and 1,628 (61.5%) were positive for IgG antibody to hepatitis B core antigen. Markers for HDV were found in 47 cases (1.7%). Antibodies to HDV were associated with Amerindian ethnic origin, a lower educational level, a history of acute viral hepatitis, a history of malaria, male sex, a history of tattooing, and older age. The most frequent HBV genotypes were A and F. This study showed a high prevalence of HBV and HDV in the western Brazilian Amazon, as well as the predominance of HBV genotypes A and F.