Monitoring human cytomegalovirus viral load in peripheral blood leukocytes of renal transplant recipients by a simple limiting dilution-PCR assay.

To assess the clinical relevance of a semi-quantitative measurement of human cytomegalovirus (HCMV) DNA in renal transplant recipients within the typical clinical context of a developing country where virtually 100% of both receptors and donors are seropositive for this virus, we have undertaken HCMV DNA quantification using a simple, semi-quantitative, limiting dilution polymerase chain reaction (PCR). We evaluated this assay prospectively in 52 renal transplant patients from whom a total of 495 serial blood samples were collected. The samples scored HCMV positive by qualitative PCR had the levels of HCMV DNA determined by end-point dilution-PCR. All patients were HCMV DNA positive during the monitoring period and a diagnosis of symptomatic infection was made for 4 of 52 patients. In symptomatic patients the geometric mean of the highest level of HCMV DNAemia was 152,000 copies per 10(6) leukocytes, while for the asymptomatic group this value was 12,050. Symptomatic patients showed high, protracted HCMV DNA levels, whereas asymptomatic patients demonstrated intermittent low or moderate levels. Using a cut-off value of 100,000 copies per 10(6) leukocytes, the limiting dilution assay had sensitivity of 100%, specificity of 92%, a positive predictive value of 43% and a negative predictive value of 100% for HCMV disease. In this patient group, there was universal HCMV infection but relatively infrequent symptomatic HCMV disease. The two patient groups were readily distinguished by monitoring with the limiting dilution assay, an extremely simple technology immediately applicable in any clinical laboratory with PCR capability.

Monitoring human cytomegalovirus viral load in peripheral blood leukocytes of renal transplant recipients by a simple limiting dilution-PCR assay

To assess the clinical relevance of a semi-quantitative measurement of human cytomegalovirus (HCMV) DNA in renal transplant recipients within the typical clinical context of a developing country where virtually 100 per cent of both receptors and donors are seropositive for this virus, we have undertaken HCMV DNA quantification using a simple, semi-quantitative, limiting dilution polymerase chain reaction (PCR). We evaluated this assay prospectively in 52 renal transplant patients from whom a total of 495 serial blood samples were collected. The samples scored HCMV positive by qualitative PCR had the levels of HCMV DNA determined by end-point dilution-PCR. All patients were HCMV DNA positive during the monitoring period and a diagnosis of symptomatic infection was made for 4 of 52 patients. In symptomatic patients the geometric mean of the highest level of HCMV DNAemia was 152,000 copies per 106 leukocytes, while for the asymptomatic group this value was 12,050. Symptomatic patients showed high, protracted HCMV DNA levels, whereas asymptomatic patients demonstrated intermittent low or moderate levels. Using a cut-off value of 100,000 copies per 106 leukocytes, the limiting dilution assay had sensitivity of 100 per cent, specificity of 92 per cent, a positive predictive value of 43 per cent and a negative predictive value of 100 per cent for HCMV disease. In this patient group, there was universal HCMV infection but relatively infrequent symptomatic HCMV disease. The two patient groups were readily distinguished by monitoring with the limiting dilution assay, an extremely simple technology immediately applicable in any clinical laboratory with PCR capability.

Highly sensitive single-step PCR protocol for diagnosis and monitoring of human cytomegalovirus infection in renal transplant recipients.

A multiplex, single-step PCR protocol for the detection of human cytomegalovirus (HCMV) DNA is described. The protocol amplifies regions of the viral LA and IE genes and employs elevated temperatures for both reagent mixing and primer annealing together with product detection by silver staining on polyacrylamide gels. This assay detects one to five HCMV genomes in clinical samples containing up to 100 ng of human DNA, a level of sensitivity equivalent to that of more complex assays involving either nested PCR or postamplification hybridization. As well as being of importance in clinical situations where high-sensitivity qualitative diagnosis is required, this assay is also applicable to the monitoring of HCMV infection in renal transplant recipients. Due to its multiplex format the assay provides quantitative information, in that samples from which a single target is amplified contain on average sevenfold fewer viral genomes per 10(6) leukocytes than those from which both targets are amplified. When weekly blood leukocyte DNA preparations from renal transplant patients were assayed, findings of three consecutive tests in which both HCMV targets were amplified were highly indicative of patients who had developed very high loads of HCMV (100% sensitivity, 88% specificity). We thus show that the same simple PCR assay which permits highly sensitive HCMV diagnosis can also be used for the efficient identification of transplant recipients at risk of clinically significant infection.

Hepatopatia em receptores de transplante renal. / Hepatopathy in renal transplantation recipients

Treze dentre 55 pacientes em hemodialise (23,6%) foram submetidos a biopsia hepaticas antes e apos transplante renal e acompanhados, em media, por 18,3 meses, por serem HBaAg positivo (12 pacientes, 21,8%) e/ou terem disfuncao hepatica. A simples persistencia do HBsAg no soro se acompanhava, com frequencia, de biopsias normais ou com alteracoes inespecificas. Entretanto, quatro pacientes HBsAg positivos apresentaram hepatopatia fatal algum tempo apos o transplante: dois com cirrose comprovada, um com necrose submacica e outro, que se tornara etilista inveterado, com provavel cirrose. Um quinto paciente HBaAg positivo evoluiu para hepatite cronica ativa apos rejeicao do transplante. Embora a persistencia do HBsAg no soro indique sempre mau prognostico apos o transplante, parece ser a principal causa de hepatopatia em nosso grupo de renais cronicos e associa-se, por vezes, a disfuncao hepatica intensa e ate fatal

Retinite por virus citomegalico. Relato de um caso. / Renitis due to cytomegalovirus: a case report.

E apresentado 1 caso de retinite por virus citomegalico (CMV), que se desenvolveu em um paciente submetido a transplante renal, em uso prolongado de terapia imunossupressiva. Os autores descrevem as caracteristicas da lesao e meios diagnosticos disponiveis, e chamam a atencao para a necessidade de um controle oftalmologico dos pacientes em uso prolongado de terapia imunossupressiva.

Tuberculose pos-transplante renal. / Tuberculosis post-renal transplantation

Em 166 pacientes transplantados de rim, 4 casos de tuberculose foram constatados, uma incidencia de 2,4% bem superior a da populacao geral brasileira de 0,3%. Tres casos foram da forma extrapulmonar, surgida entre o 2o.e 4o.anos apos transplante, e um caso de forma pulmonar, surgida um mes apos cirurgia.O quadro observado consistiu de febre, tosse, anorexia, perda de peso hepatosplenomegalia, ascite e pericardite. Todos os pacientes apresentaram PPD nao reator, leucopenia e alteracoes das provas de funcao hepatica.O diagnostico foi feito atraves de biopsia hepatica em um caso e com tratamento de prova nos tres restantes. Um paciente faleceu em decorrencia a doenca

Comportamento da hipertensao no transplantado renal com o uso oral de inibidor do enzima de conversao de angiotensina. / Behavior of hypertension in renal transplantation by oral use of an angiotensin converting enzyme inhibitor

O papel do sistema da renina como fator causador de hipertensao foi investigado em 8 pacientes submetidos a transplante renal pela administracao de captopril.Os resultados permitem propor que mais da metade das hipertensoes respondem a inibicao de formacao A II.Nao se observou absoluta correlacao entre os niveis de ARP e a resposta ao teste com captopril. Nao ha relacao entre dose de prednisona e niveis pressoricos. Pacientes com hipertensao grave tem reducao de funcao renal