RESUMO
It has been shown that the Hepatitis C virus nonstructural NS3 protein possesses at least two enzymatic domains: a serine-protease domain and an adenosine triphosphatase (ATPase)/helicase domain. In this report, a truncated fragment of NS3 (26 kDa), representing main epitopes from the (ATPase)/helicase domain, has been expressed in Escherichia coli. The recombinant protein was purified by Ion Metal Affinity Chromatography (IMAC) with more than 90% purity. The recognition of B-cell linear epitopes in the NS3 protein was evaluated by immunoblot. The recombinant NS3 protein was reduced and carboxymethylated, and the recognition of either conformational and/or linear B-cell determinants was evaluated by ELISA. The inclusion of the recombinant NS3 protein in a third-generation diagnostic system UltraMicroELISA (UMELISA) allowed an increase in the sensitivity, due to the detection of a new variety of false-negative sera in blood donor test samples.
Assuntos
Adenosina Trifosfatases/química , DNA Helicases/química , Proteínas não Estruturais Virais/química , Proteínas não Estruturais Virais/genética , Cromatografia de Afinidade , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Epitopos , Escherichia coli/metabolismo , Fermentação , Immunoblotting , Modelos Genéticos , Plasmídeos/metabolismo , Estrutura Terciária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , TemperaturaRESUMO
Infections from herpes virus play a key role in post-transplantation pathology, so it is indispensable to characterize the group of would-be renal transplant recipients. IgG antibody titers were detected by indirect immunofluorescence to Herpes simplex virus, Epstein Barr virus (EBV) and cytomegalovirus (CMV) in 100 patients treated with hemodialysis in the Nephrology Institute. The prevalence of antibodies to cytomegalovirus and to Herpes simplex and Epstein Barr viruses was 100% and 95% respectively. Average geometric titers (AGT) of antibodies to CMV were significantly higher. There was no correlation between AGT and sex, age, time of disease remission, number of blood transfusions and number of transplants.
