Estudo comparativo entre sorologia, antigenemia e reação em cadeia da polimerase para o monitoramento da infecção por citomegalovírus em pacientes receptores de transplantes de células progenitoras hematopoéticas

Forty-six allogeneic hematopoietic stem cell transplantation (HSCT) patients were monitored for the presence of CMV antibodies, CMV-DNA and CMV antigens after transplantation. Immunoenzymatic serological tests were used to detect IgM and the increase in CMV IgG antibodies (increase IgG), a nested polymerase chain reaction (N-PCR) was used to detect CMV-DNA, and an antigenemia assay (AGM) was used to detect CMV antigens. The presence of CMV-IgM and/or CMV-increase IgG antibodies was detected in 12/46 (26.1%) patients, with a median time between HSCT and the detection of positive serology of 81.5 days. A positive AGM was detected in 24/46 (52.2%) patients, with a median time between HSCT and antigen detection of 62 days. Two or more consecutive positive N-PCR results were detected in 32/46 (69.5%) patients, with a median time between HSCT and the first positive PCR of 50.5 days. These results confirmed that AGM and mainly PCR are superior to serology for the early diagnosis of CMV infection. Six patients had CMV-IgM and/or CMV-increase IgG with a negative AGM (five cases) or N-PCR assay (one case). In five of these cases the serological markers were detected during the first 100 days after HSCT, the period of highest risk. These findings support the idea that serology may be useful for monitoring CMV infections in HSCT patients, especially when PCR is unavailable.Quarenta e seis pacientes receptores de transplantes de células progenitoras hematopoéticas (TCPH) foram monitorados em relação à infecção ativa por citomegalovírus (CMV). Testes sorológicos imunoenzimáticos foram utilizados para a detecção de anticorpos IgM e elevação significativa das concentrações de anticorpos IgG (aumento IgG), nested-PCR (N-PCR) foi utilizada para a detecção de CMV-DNA e antigenemia (AGM) para a detecção de antígenos virais. A presença de CMV-IgM e/ou CMV-aumento IgG foi detectada em 12/46 (26,1%) pacientes, sendo o tempo mediano entre o transplante e a detecção dos marcadores sorológicos de 81,5 dias; AGM positiva foi detectada em 24/46 (52,2%) pacientes, sendo o tempo mediano entre o transplante e a detecção de antígenos virais de 62 dias. Dois ou mais resultados positivos consecutivos de N-PCR foram detectados em 32/46 (69,5%) pacientes, sendo o tempo mediano entre o transplante e o primeiro teste positivo de 50,5 dias. Esses resultados confirmaram que a AGM e principalmente a PCR são superiores à sorologia, com relação ao diagnóstico da infecção pelo CMV. Seis pacientes apresentaram reações CMV-IgM positivas e/ou CMV-aumento IgG com reações negativas de AGM (cinco casos) ou N-PCR (um caso). Em cinco desses casos, os marcadores sorológicos foram detectados nos 100 primeiros dias após o transplante, considerado o período de maior risco. Esses resultados indicam que os testes sorológicos podem ser úteis no monitoramento da infecção por CMV após o transplante de células progenitoras hematopoéticas, principalmente quando a N-PCR não for disponível

Active human Cytomegalovirus infection and Glycoprotein B genotypes in Brazilian pediatric renal or hematopoietic stem cell transplantation patients

A prospective analysis of active Human Cytomegalovirus infection (HCMV) was conducted on 33 pediatric renal or hematopoietic stem cell post-transplant patients. The HCMV-DNA positive samples were evaluated for the prevalence of different gB subtypes and their subsequent correlation with clinical signs. The surveillance of HCMV active infection was based on the monitoring of antigenemia (AGM) and on a nested polymerase chain reaction (N-PCR) for the detection of HCMV in the patients studied. Using restriction analysis of the gB gene sequence by PCR-RFLP (Restriction Fragment Length Polymorphism), different HCMV strains could be detected and classified in at least four HCMV genotypes. Thirty-three pediatric recipients of renal or bone marrow transplantation were monitored. Twenty out of thirty-three (60.6%) patients demonstrated active HCMV infection. gB1 and gB2 genotypes were more frequent in this population. In this study, we observed that gB2 had correlation with reactivation of HCMV infection and that patients with mixture of genotypes did not show any symptoms of HCMV disease. Future studies has been made to confirm this.505

Interfacing the operational storm surge model to a new mesoscale atmospheric mode

Chagas disease (Cd) is the third most common parasitic disease that causes damage to human health. Even a century after its description by Carlos Chagas and advances in its control, it remains a neglected disease. To eradicate the parasite or reduce the parasitic load, specific treatment for Trypanosoma cruzi (T. cruzi) is advisable; benznidazole (BNZ) is the drug that is currently prescribed. The purpose of this study is to report the adverse events (AE) due to the use of BNZ as a specific treatment for Cd, with a particular focus on hepatic changes. This was an observational, cross-sectional cohort study that included patients who were treated with BNZ. The medical records of patients who joined the Grupo de Estudo em doença de Chagas [Chagas Disease Study Group]/UNICAMP/Brazil and were treated with BNZ were reviewed for epidemiological, clinical, laboratory and AE parameters for the drug. The 204 patients who were assessed had an average age of 40.6 years ± 13.5 years, and 104 of them were women (50.98%). Fourteen (6.86%) individuals were in the acute phase of Cd, and 190 (93.13%) were in its chronic phase. AEs occurred in 85 patients (41.66%), 35 (41.17%) of whom had AEs related to the liver, characterized by an elevation of AST liver enzymes, ALT, alkaline phosphatase and gamma-glutamyltransferase (γGT). Other AEs that were observed included the following: 48 cases of cutaneous changes (56.47%), 8 cases of epigastric pain (9.41%), 7 cases of blood alteration (8.23%), and 3 cases of peripheral neuropathy (3.52%). Treatment was interrupted in 32 patients (37.64%) due to AD. Adverse events related to the liver secondary to the use of BNZ for Cd-specific treatment were frequent in this study and were characterized by an elevation of liver enzymes. Therefore, it is suggested that these enzymes be monitored during treatment with benznidazole

Retirada da terapia de manutenção para retinite por citomegalovírus em pacientes com aids exibindo resposta imunológica à terapia anti-retroviral altamente efetiva (HAART)

BACKGROUND: Before the introduction of highly active antiretroviral therapy (HAART), CMV retinitis was a common complication in patients with advanced HIV disease and the therapy was well established; it consisted of an induction phase to control the infection with ganciclovir, followed by a lifelong maintenance phase to avoid or delay relapses. METHODS: To determine the safety of CMV maintenance therapy withdrawal in patients with immune recovery after HAART, 35 patients with treated CMV retinitis, on maintenance therapy, with CD4+ cell count greater than 100 cells/mm³ for at least three months, but almost all patients presented these values for more than six months and viral load < 30000 copies/mL, were prospectively evaluated for the recurrence of CMV disease. Maintenance therapy was withdrawal at inclusion, and patients were monitored for at least 48 weeks by clinical and ophthalmologic evaluations, and by determination of CMV viremia markers (antigenemia-pp65), CD4+/CD8+ counts and plasma HIV RNA levels. Lymphoproliferative assays were performed on 26/35 patients. RESULTS: From 35 patients included, only one had confirmed reactivation of CMV retinitis, at day 120 of follow-up. No patient returned positive antigenemia tests. No correlation between lymphoproliferative assays and CD4+ counts was observed. CONCLUSION: CMV retinitis maintenance therapy discontinuation is safe for those patients with quantitative immune recovery after HAART.Antes da introdução da terapia anti-retroviral altamente efetiva (HAART), a retinite por CMV era uma complicação comum em pacientes com doença por HIV avançada e a terapia era bem estabelecida e consistia em uma fase de indução com ganciclovir para controlar a infecção, seguida por uma manutenção por toda a vida, para evitar e retardar as recidivas. Para determinar a segurança da retirada da terapia de manutenção para retinite por citomegalovírus em pacientes com recuperação imunológica após o HAART, 35 pacientes com retinite por CMV tratados com terapia de manutenção, com contagem de células CD4+ maiores que 100 células/mm³ por no mínimo três meses, mas a maioria dos pacientes apresentava esses valores por mais de seis meses e carga viral < 30.000 cópias/mL, foram avaliados prospectivamente para a recorrência de doença por CMV. A terapia de manutenção foi retirada na inclusão e os pacientes foram monitorados no mínimo 48 semanas por avaliações clínicas e oftalmológicas e pela determinação de marcadores de viremia para CMV (antigenemia). Contagens de CD4+ e CD8+ e níveis de RNA de HIV no plasma. Métodos linfoproliferativos foram realizados em 26/35 pacientes. RESULTADOS: Dos 35 pacientes incluídos no estudo, somente um teve reativação da retinite por CMV confirmada, no dia 120 do seguimento. Nenhum paciente teve testes de antigenemia positivos. Nenhuma correlação entre os ensaios linfoproliferativos e contagens de CD4+ foi observada. CONCLUSÃO: Descontinuação da terapia de manutenção para retinite por CMV é segura para aqueles pacientes com recuperação imune quantitativa após HAART

Simultaneous monitoring of CMV and human herpesvirus 6 infections and diseases in liver transplant patients: one-year follow-up

OBJECTIVE: The aim of this study was to simultaneously monitoring cytomegalovirus and human herpesvirus 6 active infections using nested-polymerase chain reaction and, together with clinical findings, follow the clinical status of patients undergoing liver transplant. INTRODUCTION: The human &#946;-herpesviruses, including cytomegalovirus and human herpesvirus 6, are ubiquitous among human populations. Active infections of human herpesvirus 6 and cytomegalovirus are common after liver transplantation, possibly induced and facilitated by allograft rejection and immunosuppressive therapy. Both viruses affect the success of the transplant procedure. METHODS: Thirty patients submitted to liver transplant at the Liver Transplant Unit, at the Gastro Center, State University of Campinas, SP, Brazil, were studied prospectively from six months to one year, nested-polymerase chain reaction for cytomegalovirus and human herpesvirus 6 DNA detections. Two or more consecutive positive nested-polymerase chain reaction were considered indicative of active infection. RESULTS: Active infection by cytomegalovirus was detected in 13/30 (43.3%) patients, median time to first cytomegalovirus detection was 29 days after transplantation (range: 0-99 days). Active infection by human herpesvirus 6 was detected in 12/30 (40%) patients, median time to first human herpesvirus 6 detection was 23.5 days after transplantation (range: 0-273 days). The time-related appearance of each virus was not statistically different (p = 0.49). Rejection of the transplanted liver was observed in 16.7% (5/30) of the patients. The present analysis showed that human herpesvirus 6 and/or cytomegalovirus active infections were frequent in liver transplant recipients at our center. CONCLUSIONS: Few patients remain free of betaherpesviruses after liver transplantation. Most patients presenting active infection with more than one virus were infected sequentially and not concurrently. Nested-polymerase chain reaction can be considered of limited value for clinically monitoring cytomegalovirus and human herpesvirus 6

Simultaneous Monitoring Of Cmv And Human Herpesvirus 6 Infections And Diseases In Liver Transplant Patients: One-year Follow-up.

The aim of this study was to simultaneously monitoring cytomegalovirus and human herpesvirus 6 active infections using nested-polymerase chain reaction and, together with clinical findings, follow the clinical status of patients undergoing liver transplant. The human β-herpesviruses, including cytomegalovirus and human herpesvirus 6, are ubiquitous among human populations. Active infections of human herpesvirus 6 and cytomegalovirus are common after liver transplantation, possibly induced and facilitated by allograft rejection and immunosuppressive therapy. Both viruses affect the success of the transplant procedure. Thirty patients submitted to liver transplant at the Liver Transplant Unit, at the Gastro Center, State University of Campinas, SP, Brazil, were studied prospectively from six months to one year, nested-polymerase chain reaction for cytomegalovirus and human herpesvirus 6 DNA detections. Two or more consecutive positive nested-polymerase chain reaction were considered indicative of active infection. Active infection by cytomegalovirus was detected in 13/30 (43.3%) patients, median time to first cytomegalovirus detection was 29 days after transplantation (range: 0-99 days). Active infection by human herpesvirus 6 was detected in 12/30 (40%) patients, median time to first human herpesvirus 6 detection was 23.5 days after transplantation (range: 0-273 days). The time-related appearance of each virus was not statistically different (p = 0.49). Rejection of the transplanted liver was observed in 16.7% (5/30) of the patients. The present analysis showed that human herpesvirus 6 and/or cytomegalovirus active infections were frequent in liver transplant recipients at our center. Few patients remain free of betaherpesviruses after liver transplantation. Most patients presenting active infection with more than one virus were infected sequentially and not concurrently. Nested-polymerase chain reaction can be considered of limited value for clinically monitoring cytomegalovirus and human herpesvirus 6.66949-5

Withdrawal of maintenance therapy for cytomegalovirus retinitis in AIDS patients exhibiting immunological response to HAART

BACKGROUND: Before the introduction of highly active antiretroviral therapy (HAART), CMV retinitis was a common complication in patients with advanced HIV disease and the therapy was well established; it consisted of an induction phase to control the infection with ganciclovir, followed by a lifelong maintenance phase to avoid or delay relapses. METHODS: To determine the safety of CMV maintenance therapy withdrawal in patients with immune recovery after HAART, 35 patients with treated CMV retinitis, on maintenance therapy, with CD4+ cell count greater than 100 cells/mm³ for at least three months, but almost all patients presented these values for more than six months and viral load < 30000 copies/mL, were prospectively evaluated for the recurrence of CMV disease. Maintenance therapy was withdrawal at inclusion, and patients were monitored for at least 48 weeks by clinical and ophthalmologic evaluations, and by determination of CMV viremia markers (antigenemia-pp65), CD4+/CD8+ counts and plasma HIV RNA levels. Lymphoproliferative assays were performed on 26/35 patients. RESULTS: From 35 patients included, only one had confirmed reactivation of CMV retinitis, at day 120 of follow-up. No patient returned positive antigenemia tests. No correlation between lymphoproliferative assays and CD4+ counts was observed. CONCLUSION: CMV retinitis maintenance therapy discontinuation is safe for those patients with quantitative immune recovery after HAART

Citomegalovirus em transplantados renais : diagnostico pela reação em cadeia da polimerase (PCR) e impacto clinico

Orientadores: Luiz Tadeu Moraes Figueiredo, Fernando Ferreira CostaTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias MedicasResumo: As complicações devidas à infecção por citomegalovirus representam um dos maiores problemas infecciosos observados em pacientes submetidos a transplantes em geral. Com o objetivo de avaliar a incidência e o impacto clínico da infecção por CMV em uma população de transplantados renais no Brasil foram estudados prospectivamente 37 pacientes submetidos a transplante renal no Hospital das Clínicas da UNICAMP, no período de Outubro de 1990 a Setembro de 1992. A identificação da infecção pelo CMV foi levada a efeito pela detecção direta de partículas virais na urina através da reação em cadeia da polimerase (PCR) e por reações soro lógicas - ELISA e imunofluorescência indireta. A PCR foi realizada com 3 pares de iniciadores ("primers"). O material amplificado foi fixado em filtros de nylon como "dot-blot" e as partículas virais foram identificadas por hibridização com oligonucleotídeos específicos para a região amplificada (sonda), marcados com 32P por meio da T4 polinucleotídeo quinase. O material para análise foi obtido antes e em intervalos mensais após o transplante. Os resultados globais, agrupando sorologia e PCR mostraram 32 pacientes com evidências de infecção ativa pelo CMV entre 37 estudados (86,48%). Em 6 pacientes, só a PCR foi capaz de identificar infecção ativa. O diagnóstico foi mais precoce pela PCR do que pelas técnicas sorológicas. 10 pacientes já apresentavam virúria antes do transplante. A maior incidência de infecção ocorreu nos primeiros 4 meses que se seguiram ao transplante. Dos 32 pacientes com infecção ativa pelo CMV, 16 (50%) apresentaram manifestações clinicas sugestivas de serem secundária à replicação viral (doença por CMV). Os 5 pacientes que não apresentaram, por nenhum dos testes, evidências de infecção ativa pelo CMV, não tiveram problemas clínicos relevantes no seguimento e mantiveram boa função renal. Em seu conjunto, nossos resultados sugerem que a infecção pelo CMV é uma complicação frequente em transplantados renais no Brasil e que poderia estar associada à maior frequência de complicações clínicas nesses pacientes. Adicionalmente, comprovou-se que a PCR é um método eficaz de detecção precoce dessa infecção.Abstract: Cytomegalovirus (C.M.V.) is the single most important infections agent affecting recipients of organ transplant. To evaluate the incidence and the clinical impact of cytomegalovirus infection on renal transplant in Brazil, 37 patients who underwent renal allograft transplants were periodically screened for the presence of C.M.V. virus DNA in urine by lIsing polymerase chain reaction (P.C.R.) and for the presence of IgM and IgG cytomcgalovirlls antibodies (ELlSA and I.F.I.). The P.C.R. amplificd prodllcts werc detected by gel electrophoresis and by dot blot hybridization with oligonucleotide probes. 32 out 37 patients (86.48%) were found to be positive by at least one of the three methods. In 6 patients, P.C.R. was the only test to detect the C.M.V. infection. 10 patients had a positive result by P.C.R.Qefore the transplantation. The diagnosis was made earlier by P.C.R. than by serologic tests. Active infection occured more frequently in the first 4 months after transplantation. 16 out 32 patients (50.00%) with active infection by C.M.V. presented clinical manifestations of C.M. V. disease. 5 patients without evidences of active infection by the 3 tests have only minor clinical complications during followup. Our results sugest that C.M.V. infection is a frequent problem in renal transplant patients in Brazil, with significant clinical complications. Moreover, we confirmed that P.C.R. is a very sensitive procedure to early detection of C.M.V. infection.DoutoradoDoutor em Clínica Médic