Diagnóstico da hepatite por vírus C em doadores de sangue brasileiros, usando a reação de transcrição reversa e a reação em cadeia da polimerase "nested": comparação com os ensaios imunoenzimáticos e imunoblot recombinante

Screening blood donations for anti-HCV antibodies and alanine aminotransferase (ALT) serum levels generally prevents the transmission of hepatitis C virus (HCV) by transfusion. The aim of the present study was to evaluate the efficiency of the enzyme immunoassay (EIA) screening policy in identifying potentially infectious blood donors capable to transmit hepatitis C through blood transfusion. We have used a reverse transcriptase (RT)-nested polymerase chain reaction (PCR) to investigate the presence of HCV-RNA in blood donors. The prevalence of HCV-RNA positive individuals was compared with the recombinant immunoblot assay (RIBA-2) results in order to assess the usefulness of both tests as confirmatory assays. Both tests results were also compared with the EIA-2 OD/C ratio (optical densities of the samples divided by the cut off value). ALT results were expressed as the ALT quotient (qALT), calculated dividing the ALT value of the samples by the maximum normal value (53UI/l) for the method. Donors (n=178) were divided into five groups according to their EIA anti-HCV status and qALT: group A (EIA >; or = 3, ALT; or = 3, ALT>;1), group C (1Na prevenção da transmissão de Hepatite por Vírus C (HCV) em transfusões de hemocomponentes, utiliza-se rotineiramente, como testes de triagem de doadores de sangue, ensaios que detectam anticorpos anti-HCV e dosagens da enzima alanina-aminotransferase (ALT). O presente estudo tem como objetivo principal avaliar a eficiência do ensaio imunoenzimático de segunda geração (EIA-2) como teste de triagem, na identificação de doadores de sangue potencialmente infectados, e portanto, capazes de transmitir hepatite C pelos hemocomponentes. Nós utilizamos o ensaio de transcrição reversa (RT) e a reação em cadeia da polimerase "nested" («nested PCR») para investigar a presença do RNA do vírus da hepatite C (HCV) em doadores de sangue. A prevalência do RNA-HCV em indivíduos positivos foi comparada com os resultados do ensaio complementar imunoblot recombinante de segunda geração (RIBA-2) com o intuito de avaliar a utilidade de ambos como testes confirmatórios. Estes dois testes também foram comparados com a razão DO/C (valores de densidade óptica das amostras dividida pelo valor de corte da reação) no EIA-2. Os resultados das dosagens da ALT foram expressos como uma razão unitária denominada qALT, que representa o cálculo do valor do ALT da amostra dividido pelo valor máximo considerado normal para o teste (53UI/L). Os doadores de sangue foram divididos em cinco grupos de acordo com os resultados do EIA-2 e o qALT: grupo A (EIA >; ou = 3, ALT; ou = 3, ALT>;1), grupo C (

Hepatitis C virus in monozygotic twins

É relatado o caso de paciente grávida, com hepatite C crônica que deu à luz dois gêmeos monozigóticos. Um recém-nascido apresentou positividade para o RNA do vírus da hepatite C (RNA-VHC), no sangue venoso, coletado de veia periférica doze horas após o parto. O outro recém-nascido apresentou-se negativo para o RNA-VHC logo após o nascimento, porém tornou-se RNA-VHC positivo na amostra coletada aos três meses de idade. Os resultados permitem supor que um dos gêmeos provavelmente foi contaminado no período intra-uterino, enquanto o outro adquiriu a infecção no período perinatal. Ambos foram negativos para a presença do RNA-VHC e para os anticorpos anti-HCV em todas as amostras séricas coletadas após os nove meses de idade. Os exames laboratoriais dos gêmeos não mostraram a presença de infecção crônica pelo VHC durante o acompanhamento de 29 meses .A case of a pregnant patient with chronic hepatitis C who gave birth to monozygotic twins that were infected with HCV is reported. One of the newborns was positive for HCV-RNA in blood sample collected 12 hours after delivery. The other newborn was negative for HCV-RNA at birth, but was detected HCV viremia at three months of age. The results have led to the conclusion that one of the twins was probably contaminated in the intrauterine period, while the other acquired the infection in the perinatal period. Both were negative for HCV-RNA and for anti-HCV in the serum samples collected at nine months of age. The report describes the changes in the laboratory tests conducted in mother and twins until 29 months after delivery

Extended treatment with interferon and ribavirin in a hemodialysis patient with chronic hepatitis C

Hemodialysis patient with chronic HCV infection,who was started on monotherapy with interferon.Qualitative HCV RNA remained positive at 12 weeks of treatment; ribavirin was associated. HCV RNA was negative at week 24 and treatment was extended to 72 weeks. HCV RNA negative six months after treatment.19319

Avaliação clínica, epidemiológica, laboratorial, histológica e ultrassonográfica de doadores de sangue anti-HCV EIA-2 positivos

Between 1992 and 1997, 790 blood donors with anti-HCV EIA-2 strongly reagent (relationship between the sample optical density/cut-off >; 3) detected at the blood bank serological screening, were evaluated in ambulatory environment. They were all negative for Chagas disease, syphilis, hepatitis B (HBsAg) and AIDS. Blood samples were collected at the first ambulatorial evaluation, for hemogram, biochemical tests and new serological tests for HCV (anti-HCV EIA-2). In blood samples of 226 repeatedly reagent anti-HCV EIA-2 blood donors, supplementary "immunoblot" test for HCV (RIBA-2) was used. In 209 donors, the presence of HCV-RNA was investigated by the PCR test. The abdominal ultrasonography was realized in 366 donors. In 269 patients liver biopsy was performed for the histopathological study. The follow-up of blood donors showed that 95.6% were repeatedly EIA-2 reagent, 94% were symptomless and denied any hepatitis history, with only 2% mentioning previous jaundice. In 47% of this population at least one risk factor has been detected for the HCV transmission, the use of intravenous drugs being the main one (27.8%). Blood transfusion was the second factor for HCV transmission (27.2%). Hepatomegaly was detected in 54% of the cases. Splenomegaly and signs of portal hypertension have seldom been found in the physical examination, indicating a low degree of hepatic compromising in HCV. Abdominal ultrasound showed alterations in 65% of the subjects, being the steatosis the most frequent (50%). In 83.5% of the donors submitted to the liver biopsy, the histopathological exam showed the presence of chronic hepatitis, usually classified as active (89%) with mild or moderate grade in most of the cases (99.5%). The histopathological exam of the liver was normal in 1.5% of blood donors. The RIBA-2 test and the HCV-RNA investigation by PCR were positive in respectively 91.6 and 75% of the anti-HCV EIA-2 reagent donors. The HCV-RNA research was positive in 82% of the RIBA-2 positive subjects, in 37.5% of the indeterminate RIBA-2 donors and in 9% of the negative RIBA-2 donors. Chronic hepatitis has also been observed in 50% of the histopathological exams of the anti-HCV EIA-2 reagent donors which were indeterminate RIBA-2. Among 18 blood donors with minimal changes histopathological exam 11 (61%) were HCV-RNA positive. Our blood donors anti-HCV reagent generally had clinical, laboratorial and histopathological features observed in patients with chronic HCV hepatitis and a high proportion could be identified in interviews and medical evaluation realized in blood blanks. Generally, these HCV infected donors are identified and discharged only by the serological tests results.Entre 1992 e 1997 foram avaliados, ambulatorialmente, 790 doadores de sangue com teste anti-HCV EIA-2 fortemente reagente (relação entre a densidade ótica da amostra / "cut-off" >; 3), que haviam sido detectados na triagem sorológica do banco de sangue. Todos eram negativos para doença de Chagas, sífilis, hepatite B (HBsAg) e AIDS. Amostras de sangue foram coletadas, na primeira consulta ambulatorial, para a realização de hemograma, exames bioquímicos e novos testes sorológicos para a HVC (anti-HCV EIA-2). Em 226 doadores anti-HCV EIA-2 repetidamente reagentes, realizou-se o teste suplementar de "immunoblot" para a HVC (RIBA-2). Em 209 doadores, pesquisou-se a presença do RNA do VHC pelo teste do PCR, através de exame automatizado (HCV-AMPLICOR, ROCHE). A ultra-sonografia abdominal foi realizada em 366 doadores e a biópsia hepática em 269 concordantes. Notou-se que 95,6% eram EIA-2 repetidamente reagentes, 94% eram assintomáticos e que apenas 2% referiram icterícia pregressa. Em 47% detectou-se, pelo menos, um fator de risco para a transmissão do VHC, sendo o uso de drogas E.V. o principal deles (27,8%). A transfusão de sangue foi o segundo fator na transmissão da HVC (27,2%). Hepatomegalia foi encontrada em 54%. Esplenomegalia e sinais de hipertensão portal foram raramente encontrados no exame físico, denotando o baixo grau de comprometimento hepático na HVC. A ultra-sonografia abdominal mostrou-se alterada em 65% dos indivíduos, sendo a esteatose a alteração mais freqüentemente observada (50%). Em 83,5% dos doadores submetidos à biópsia hepática, diagnosticou-se hepatite crônica, geralmente classificada como ativa (89%) e de grau leve ou moderado na maioria dos casos (99,5%). O histopatológico foi normal em 1,5% dos doadores. O teste de RIBA-2 e a pesquisa do RNA do VHC pelo PCR foram positivos em, respectivamente, 91,6 e 75% dos doadores anti-HCV EIA-2 reagentes. A pesquisa do RNA do VHC foi positiva em 82% dos indivíduos RIBA-2 reagentes, em 37,5% dos doadores RIBA-2 indeterminados e em 9% dos RIBA-2 negativos. Hepatite crônica foi observada em 50% dos doadores RIBA-2 indeterminados. Entre 18 doadores com alterações mínimas, ao exame histopatológico, 11 (61%) eram positivos para o RNA do VHC. Nossos doadores de sangue anti-HCV reagentes geralmente apresentam alterações clínicas, laboratoriais e histopatológicas próprias de pacientes com hepatites crônicas pelo VHC e uma elevada proporção destes podem ser identificados em entrevistas e avaliação médicas rotineiramente realizadas em bancos de sangue. Geralmente estes doadores infectados pelo VHC somente são identificados e bloqueados pelos resultados dos testes sorológicos

Peginterferon alfa-2a plus ribavirin for chronic hepatitis C virus infection.

BACKGROUND Treatment with peginterferon alfa-2a alone produces significantly higher sustained virologic responses than treatment with interferon alfa-2a alone in patients with chronic hepatitis C virus (HCV) infection. We compared the efficacy and safety of peginterferon alfa-2a plus ribavirin, interferon alfa-2b plus ribavirin, and peginterferon alfa-2a alone in the initial treatment of chronic hepatitis C. METHODS A total of 1121 patients were randomly assigned to treatment and received at least one dose of study medication, consisting of 180 μg of peginterferon alfa-2a once weekly plus daily ribavirin (1000 or 1200 mg, depending on body weight), weekly peginterferon alfa-2a plus daily placebo, or 3 million units of interferon alfa-2b thrice weekly plus daily ribavirin for 48 weeks. RESULTS A significantly higher proportion of patients who received peginterferon alfa-2a plus ribavirin had a sustained virologic response (defined as the absence of detectable HCV RNA 24 weeks after cessation of therapy) than of patients who received interferon alfa-2b plus ribavirin (56 percent vs. 44 percent, P CONCLUSIONS In patients with chronic hepatitis C, once-weekly peginterferon alfa-2a plus ribavirin was tolerated as well as interferon alfa-2b plus ribavirin and produced significant improvements in the rate of sustained virologic response, as compared with interferon alfa-2b plus ribavirin or peginterferon alfa-2a alone

Peginterferon Alfa-2a plus Ribavirin for Chronic Hepatitis C Virus Infection

Background Treatment with peginterferon alfa-2a alone produces significantly higher sustained virologic responses than treatment with interferon alfa-2a alone in patients with chronic hepatitis C virus (HCV) infection. We compared the efficacy and safety of peginterferon alfa-2a plus ribavirin, interferon alfa-2b plus ribavirin, and peginterferon alfa-2a alone in the initial treatment of chronic hepatitis C. Methods A total of 1121 patients were randomly assigned to treatment and received at least one dose of study medication, consisting of 180 μg of peginterferon alfa-2a once weekly plus daily ribavirin (1000 or 1200 mg, depending on body weight), weekly peginterferon alfa-2a plus daily placebo, or 3 million units of interferon alfa-2b thrice weekly plus daily ribavirin for 48 weeks. Results A significantly higher proportion of patients who received peginterferon alfa-2a plus ribavirin had a sustained virologic response (defined as the absence of detectable HCV RNA 24 weeks after cessation of therapy) than of patients who received interferon alfa-2b plus ribavirin (56 percent vs. 44 percent, P<0.001) or peginterferon alfa-2a alone (56 percent vs. 29 percent, P<0.001). The proportions of patients with HCV genotype 1 who had sustained virologic responses were 46 percent, 36 percent, and 21 percent, respectively, for the three regimens. Among patients with HCV genotype 1 and high base-line levels of HCV RNA, the proportions of those with sustained virologic responses were 41 percent, 33 percent, and 13 percent, respectively. The overall safety profiles of the three treatment regimens were similar; the incidence of influenza-like symptoms and depression was lower in the groups receiving peginterferon alfa-2a than in the group receiving interferon alfa-2b plus ribavirin. Conclusions In patients with chronic hepatitis C, once-weekly peginterferon alfa-2a plus ribavirin was tolerated as well as interferon alfa-2b plus ribavirin and produced significant improvements in the rate of sustained virologic response, as compared with interferon alfa-2b plus ribavirin or peginterferon alfa-2a alone

Diagnóstico da hepatite por vírus C em doadores de sangue brasileiros, usando a reação de transcrição reversa e a reação em cadeia da polimerase nested: comparação com os ensaios imunoenzimáticos e imunoblot recombinante

Screening blood donations for anti-HCV antibodies and alanine aminotransferase (ALT) serum levels generally prevents the transmission of hepatitis C virus (HCV) by transfusion. The aim of the present study was to evaluate the efficiency of the enzyme immunoassay (EIA) screening policy in identifying potentially infectious blood donors capable to transmit hepatitis C through blood transfusion. We have used a reverse transcriptase (RT)-nested polymerase chain reaction (PCR) to investigate the presence of HCV-RNA in blood donors. The prevalence of HCV-RNA positive individuals was compared with the recombinant immunoblot assay (RIBA-2) results in order to assess the usefulness of both tests as confirmatory assays. Both tests results were also compared with the EIA-2 OD/C ratio (optical densities of the samples divided by the cut off value). ALT results were expressed as the ALT quotient (qALT), calculated dividing the ALT value of the samples by the maximum normal value (53UI/l) for the method. Donors (n=178) were divided into five groups according to their EIA anti-HCV status and qALT: group A (EIA > or = 3, ALT or = 3, ALT>1), group C (11) and group E (EIA or =3 and detectable HCV-RNA by RT-nested PCR. We have also noted that blood donors with RIBA-2 indeterminate presented a high degree of detectable HCV-RNA using RT-nested PCR (75%), especially when the c22.3 band was detected.Na prevenção da transmissão de Hepatite por Vírus C (HCV) em transfusões de hemocomponentes, utiliza-se rotineiramente, como testes de triagem de doadores de sangue, ensaios que detectam anticorpos anti-HCV e dosagens da enzima alanina-aminotransferase (ALT). O presente estudo tem como objetivo principal avaliar a eficiência do ensaio imunoenzimático de segunda geração (EIA-2) como teste de triagem, na identificação de doadores de sangue potencialmente infectados, e portanto, capazes de transmitir hepatite C pelos hemocomponentes. Nós utilizamos o ensaio de transcrição reversa (RT) e a reação em cadeia da polimerase nested («nested PCR») para investigar a presença do RNA do vírus da hepatite C (HCV) em doadores de sangue. A prevalência do RNA-HCV em indivíduos positivos foi comparada com os resultados do ensaio complementar imunoblot recombinante de segunda geração (RIBA-2) com o intuito de avaliar a utilidade de ambos como testes confirmatórios. Estes dois testes também foram comparados com a razão DO/C (valores de densidade óptica das amostras dividida pelo valor de corte da reação) no EIA-2. Os resultados das dosagens da ALT foram expressos como uma razão unitária denominada qALT, que representa o cálculo do valor do ALT da amostra dividido pelo valor máximo considerado normal para o teste (53UI/L). Os doadores de sangue foram divididos em cinco grupos de acordo com os resultados do EIA-2 e o qALT: grupo A (EIA > ou = 3, ALT ou = 3, ALT>1), grupo C (11) e grupo E (EIA ou = 3 e a positividade do RNA-HCV por RT-nested PCR. Nós também pudemos observar que os doadores de sangue com RIBA-2 indeterminados apresentaram um alto grau de positividade no RT-nested PCR (75%), especialmente quando está presente a banda correspondente ao antígeno c22.3.26326