RESUMEN
INTRODUCTION: Human cytomegalovirus is one of the causes of opportunist infections in immunocompromised patients, and is triggered by factors such as state of viral latency, weakened immune responses, and development of antiviral resistance to ganciclovir, the only drug offered by the public health system in Brazil to treat the infection. The goal of this study was to identify mutations that may be associated with antiviral resistance in immunocompromised patients. METHODS: Molecular analysis was performed in 82 blood samples and subjected to genomic DNA extraction by a silica-based method. Three sequences of the HCMV UL97 gene, which encodes a phosphotransferase protein required for activation of ganciclovir, were amplified by polymerase chain reaction. Pyrosequencing methods were applied to one external 2096-bp segment DNA and two internal sequences between nucleotides 1087 to 1828 to detect mutations in this gene. RESULTS: Approximately 10% of sequences contained mutations between nucleotides 377 and 594, in conserved regions of the UL97 gene, leading to amino acid changes. Eleven coding mutations were identified, including changes leading to amino acid substitutions, E596K and S604F, which were observed in 100% of samples and are described for the first time in Brazil. In addition, one mutation (A594V) that is associated with ganciclovir resistance was detected in a kidney transplant patient. CONCLUSIONS: Further studies to detect mutations associated with HCMV resistance to antiviral drugs are required to demonstrate the need to increase the variety and availability of drugs used to treat viral infections in the public health care system in Brazil.
Asunto(s)
Antivirales/uso terapéutico , Infecciones por Citomegalovirus/tratamiento farmacológico , Citomegalovirus/enzimología , Farmacorresistencia Viral/genética , Huésped Inmunocomprometido , Mutación/genética , Fosfotransferasas/genética , Antivirales/farmacología , Estudios de Casos y Controles , Estudios Transversales , Citomegalovirus/efectos de los fármacos , Citomegalovirus/genética , Farmacorresistencia Viral/efectos de los fármacos , Genotipo , Humanos , Reacción en Cadena de la PolimerasaRESUMEN
Abstract INTRODUCTION: Human cytomegalovirus is one of the causes of opportunist infections in immunocompromised patients, and is triggered by factors such as state of viral latency, weakened immune responses, and development of antiviral resistance to ganciclovir, the only drug offered by the public health system in Brazil to treat the infection. The goal of this study was to identify mutations that may be associated with antiviral resistance in immunocompromised patients. METHODS: Molecular analysis was performed in 82 blood samples and subjected to genomic DNA extraction by a silica-based method. Three sequences of the HCMV UL97 gene, which encodes a phosphotransferase protein required for activation of ganciclovir, were amplified by polymerase chain reaction. Pyrosequencing methods were applied to one external 2096-bp segment DNA and two internal sequences between nucleotides 1087 to 1828 to detect mutations in this gene. RESULTS: Approximately 10% of sequences contained mutations between nucleotides 377 and 594, in conserved regions of the UL97 gene, leading to amino acid changes. Eleven coding mutations were identified, including changes leading to amino acid substitutions, E596K and S604F, which were observed in 100% of samples and are described for the first time in Brazil. In addition, one mutation (A594V) that is associated with ganciclovir resistance was detected in a kidney transplant patient. CONCLUSIONS: Further studies to detect mutations associated with HCMV resistance to antiviral drugs are required to demonstrate the need to increase the variety and availability of drugs used to treat viral infections in the public health care system in Brazil.
Asunto(s)
Humanos , Antivirales/uso terapéutico , Fosfotransferasas/genética , Huésped Inmunocomprometido , Infecciones por Citomegalovirus/tratamiento farmacológico , Citomegalovirus/enzimología , Farmacorresistencia Viral/genética , Mutación/genética , Antivirales/farmacología , Estudios de Casos y Controles , Reacción en Cadena de la Polimerasa , Estudios Transversales , Citomegalovirus/efectos de los fármacos , Citomegalovirus/genética , Farmacorresistencia Viral/efectos de los fármacos , GenotipoRESUMEN
INTRODUCTION: Human cytomegalovirus is an opportunistic betaherpesvirus that causes persistent and serious infections in immunodeficient patients. Recurrent infections occur due to the presence of the virus in a latent state in some cell types. It is possible to examine the virus using molecular methods to aid in the immunological diagnosis and to generate a molecular viral profile in immunodeficient patients. The objective of this study was to characterize cytomegalovirus genotypes and to generate the epidemiological and molecular viral profile in immunodeficient patients. METHODS: A total of 105 samples were collected from immunodeficient patients from the City of Belém, including newborns, hemodialysis patients, transplant recipients and HIV+ patients. An IgG and IgM antibody study was completed using ELISA, and enzymatic analysis by restriction fragment length polymorphism (RFLP) was performed to characterize viral genotypes. RESULTS: It was observed that 100 percent of the patients had IgG antibodies, 87 percent of which were IgG+/IgM-, consistent with a prior infection profile, 13 percent were IgG+/IgM+, suggestive of recent infection. The newborn group had the highest frequency (27 percent) of the IgG+/IgM+ profile. By RFLP analysis, only one genotype was observed, gB2, which corresponded to the standard AD169 strain. CONCLUSIONS: The presence of IgM antibodies in new borns indicates that HCMV continues to be an important cause of congenital infection. The low observed genotypic diversity could be attributed to the small sample size because newborns were excluded from the RFLP analysis. This study will be continued including samples from newborns to extend the knowledge of the general and molecular epidemiology of HCMV in immunodeficient patients.
INTRODUÇÃO: O citomegalovírus é um betaherpesvírus oportunista, causador de infecções persistentes e graves em pacientes imunodeficientes. As infecções recorrentes ocorrem devido à presença do vírus em estado de latência, em alguns tipos celulares, o que possibilita a pesquisa viral por métodos moleculares para auxiliar nos diagnósticos imunológicos, assim como traçar o perfil epidemiológico e molecular viral em pacientes imunodeficientes. O objetivo deste estudo foi caracterizar os genótipos de citomegalovírus e traçar o perfil epidemiológico e molecular viral em pacientes imunodeficientes. MÉTODOS: Um total de 105 amostras foi coletado de pacientes imunodeficientes da Cidade de Belém, incluindo recém-nascidos, hemodialisados, transplantados e pacientes HIV+. Foi realizada a pesquisa de anticorpos IgG e IgM pelo método ELISA e análise enzimática pelo método restriction fragment length polymorphism (RFLP) para caracterização dos genótipos virais. RESULTADOS: Foi observado que 100 por cento dos pacientes apresentavam anticorpos IgG, 87 por cento eram IgG+/IgM-perfil de infecção pregressa; e 13 por cento IgG+/ IgM+ sugestivo de infecção recente. O grupo dos recém-nascidos apresentou maior frequência (27 por cento) do perfil IgG+/IgM+. Na análise por RFLP, foi observado um único genótipo, o gB2, que corresponde ao padrão genotípico da cepa AD169. CONCLUSÕES: A presença de anticorpos IgM nos recém-nascidos indica que o vírus CMV continua sendo causa importante de infecção congênita; a baixa diversidade genotípica pode ser atribuída ao tamanho amostral devido a exclusão dos recém-nascidos na análise por RFLP. Esse estudo será continuado incluindo amostras de recém-nascidos a fim de contribuir para um amplo conhecimento da epidemiologia geral e molecular do citomegalovírus em pacientes imunodeficientes da Cidade de Belém.
Asunto(s)
Adulto , Humanos , Recién Nacido , Infecciones por Citomegalovirus/virología , Citomegalovirus/genética , Genoma Viral/genética , Infecciones por VIH/inmunología , Huésped Inmunocomprometido/inmunología , Trasplante de Riñón/inmunología , Brasil , Diálisis , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Mapeo Restrictivo/métodosRESUMEN
INTRODUCTION: Human cytomegalovirus is an opportunistic betaherpesvirus that causes persistent and serious infections in immunodeficient patients. Recurrent infections occur due to the presence of the virus in a latent state in some cell types. It is possible to examine the virus using molecular methods to aid in the immunological diagnosis and to generate a molecular viral profile in immunodeficient patients. The objective of this study was to characterize cytomegalovirus genotypes and to generate the epidemiological and molecular viral profile in immunodeficient patients. METHODS: A total of 105 samples were collected from immunodeficient patients from the City of Belém, including newborns, hemodialysis patients, transplant recipients and HIV+ patients. An IgG and IgM antibody study was completed using ELISA, and enzymatic analysis by restriction fragment length polymorphism (RFLP) was performed to characterize viral genotypes. RESULTS: It was observed that 100% of the patients had IgG antibodies, 87% of which were IgG+/IgM-, consistent with a prior infection profile, 13% were IgG+/IgM+, suggestive of recent infection. The newborn group had the highest frequency (27%) of the IgG+/IgM+ profile. By RFLP analysis, only one genotype was observed, gB2, which corresponded to the standard AD169 strain. CONCLUSIONS: The presence of IgM antibodies in new borns indicates that HCMV continues to be an important cause of congenital infection. The low observed genotypic diversity could be attributed to the small sample size because newborns were excluded from the RFLP analysis. This study will be continued including samples from newborns to extend the knowledge of the general and molecular epidemiology of HCMV in immunodeficient patients.
Asunto(s)
Infecciones por Citomegalovirus/virología , Citomegalovirus/genética , Genoma Viral/genética , Infecciones por VIH/inmunología , Huésped Inmunocomprometido/inmunología , Trasplante de Riñón/inmunología , Adulto , Brasil , Diálisis , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Recién Nacido , Mapeo Restrictivo/métodosRESUMEN
Descreve o perfil soroepidemiológico da Rubéola de pacientes referenciados ao Instituto Evandro Chagas/SVS/MS nos períodos pré-vacinal(1989 a 1999) e pós-vacinal (2000 a 2005), foi realizado estudo retrospectivo do banco de dados de 34.221 amostras, cujos testes sorológicos foram analisados através da técnica de pesquisa de IgM e IgG por ELISA com kits do laboratório DADE BEHRING. A taxa de infecção encontrada foi de 17,2% no período pré-vacinal e de 4,0% no pós-vacinal. Entre a sintomatologia apresentada no período pré-vacinal, a linfadenopatia teve maior taxa com 38,4% e no pós-vacinal a artralgia com 11,3%. Nas mulheres em idade fértil, a média da taxa de imunes foi de 78,3% e 84,4% no período pré e pós-vacinal, respectivamente. A taxa de infecção em gestantes no período pré-vacinal foi de 9,3% e no pós-vacinal 1,6%. Os recém-nascidos infectados corresponderam a 2,1% no período pré e 1,0% no período pós-vacinal nesses, houve predomínio de catarata e cardiopatia isoladas ou em associação. Foi concluído que houve diferença significante entre as frequências de todos os seguimentos estudados, em relação aos períodos pré e pós-vacinal, confirmando a eficácia da vacina na prevenção da Rubéola e da SRC, tal fato realça a necessidade de se ampliar as coberturas vacinais para impedir a circulação do VR no país, cumprindo assim o acordo de eliminação até o ano 2010
Asunto(s)
Masculino , Femenino , Humanos , Síndrome de Rubéola Congénita , Rubéola (Sarampión Alemán)/diagnóstico , Rubéola (Sarampión Alemán)/etiología , Rubéola (Sarampión Alemán)/transmisión , Estudios SeroepidemiológicosRESUMEN
O citomegalovírus (CMV) é uma das principais causas de morbimortalidade de pacientes imunodeficientes. Em indivíduos transplantados, a infecção pelo vírus pode levar a rejeição aguda do enxerto, devido à produção de citocinas em resposta à infecção. Foi realizado o diagnóstico laboratorial da infecção pelo CMV em 35 pacientes transplantados renais através dos métodos ELISA e PCR no sangue, sendo acompanhados 14 indivíduos nos períodos pré e pós-transplante. As análises foram efetuadas no Instituto Evandro Chagas para confirmar o diagnóstico clínico da infecção viral e descrever o perfil sorológico-molecular dos pacientes. Dos 14 indivíduos avaliados no pré-transplante, 78,5 por cento apresentavam IgG e 7,1 por cento IgM para infecção por CMV. Na sorologia dos 35 pacientes no pós-transplante obteve-se 100 por cento de positividade para IgG e 14,5 por cento para IgM. Indivíduos soronegativos no pré-transplante soroconverteram após o enxerto com presença de IgG e IgM. Na análise molecular, 17,1 por cento apresentaram DNA viral, e os perfis sorológico-molecular foram: 2,8 por cento IgGpositivo IgMpositivoPCRpositivo; 14,3 por cento IgGpositivoIgMnegativoPCRpositivo; 8,6 por cento IgGpositivoIgMpositivoPCRnegativo e 74,3 por cento IgGpositivoIgMnegativoPCRnegativo. O perfil sorológico-molecular predominante é compatível com o perfil epidemiológico de elevada circulação do vírus na população. O conjunto IgGpositivoIgMnegativoPCRpositivo sugere a possibilidade de reinfecção por reativação viral em 8,6 por cento dos pacientes transplantados que pode ter sido da cepa exógena do doador para o receptor.