RESUMO
AIM: To produce and test recombinant multiepitope proteins as an alternative assay for the serological diagnosis of cryptococcosis. MATERIALS & METHODS: Previously, synthetic peptides were used to detect anti-Cryptococcus antibodies, and in silico analyses showed that the union of peptides would improve the results. Here, the coding sequences of these peptides were assembled into synthetic genes. Four genes have been cloned and expressed in Escherichia coli, producing recombinant multiepitope proteins: proteins A, B, C and D. RESULTS: All constructs yielded good results; however, protein D showed the best results, with a sensitivity of 88.57% and specificity of 100%. CONCLUSION: The multiepitope proteins were shown to be potential antigens for the diagnosis of cryptococcosis in an attempt to detect anti-Cryptococcus antibodies.
Assuntos
Anticorpos Antifúngicos/imunologia , Criptococose/diagnóstico , Cryptococcus/imunologia , Epitopos de Linfócito B/imunologia , Proteínas Recombinantes/imunologia , Sequência de Aminoácidos , Anticorpos Antifúngicos/sangue , Antígenos de Fungos/genética , Antígenos de Fungos/imunologia , Criptococose/sangue , Criptococose/imunologia , Criptococose/microbiologia , Cryptococcus/genética , Ensaio de Imunoadsorção Enzimática , Epitopos de Linfócito B/genética , Escherichia coli/genética , Humanos , Proteínas Recombinantes/genética , Sensibilidade e EspecificidadeRESUMO
AIM: To determine the immunoreactivity of synthetic Cryptococcus-derived peptides. MATERIALS & METHODS: A total of 63 B-cell epitopes from previously identified Cryptococcus gattii immunoreactive proteins were synthesized and evaluated as antigens in ELISAs. The peptides were first evaluated for their ability to react against sera from immunocompetent subjects carrying cryptococcal meningitis. Peptides that yielded high sensitivity and specificity in the first test were then retested with sera from individuals with other fungal pathologies for cross-reactivity determination. RESULTS: Six of 63 synthetic peptides were recognized by antibodies in immunoassays, with a specificity of 100%, sensitivity of 78% and low cross-reactivity. CONCLUSION: We successfully determined the immunoreactivity of selected synthetic peptides of C. gattii derived proteins.
Assuntos
Proteínas de Bactérias/imunologia , Cryptococcus gattii/imunologia , Peptídeos/imunologia , Anticorpos Antifúngicos/análise , Anticorpos Antifúngicos/imunologia , Proteínas de Bactérias/genética , Criptococose/imunologia , Criptococose/microbiologia , Cryptococcus gattii/genética , Ensaio de Imunoadsorção Enzimática , Epitopos de Linfócito B/imunologia , Humanos , Peptídeos/síntese química , Peptídeos/químicaRESUMO
Histoplasmosis is an endemic mycosis in specific areas of the U.S. and Latin America, caused by dimorphic geophilic fungus H. capsulatum. The description of cases in Brazil's northeast, considering that this region isn't an endemic area, indicates the existence of small outbreaks in several regions. Precise reasons for this pattern of distribution are still unknown. Activities related to the removal of soil in these places are associated with exposure to H. capsulatum. The objective of this work is to describe three isolated cases of histoplasmosis which occurred in Brazil's northeast region in individuals exposed to different activities (demolition of houses, wells diggers and exposure to bats).
