Evaluación y uso de un péptido sintético para la detección de anticuerpos específicos contra Plasmodium falciparum / Development and use of a synthetic peptide for the detection of specific antibodies against Plasmodium falciparum

Resumen Introducción. La malaria continúa siendo una de las enfermedades que causa mayor morbi-mortalidad a nivel mundial. Por esta razón es importante desarrollar herramientas diagnósticas eficaces que se implementen como estrategias para el control de la enfermedad. Objetivos. Estandarizar las condiciones del inmunoensayo enzimático (ELISA), para la detección de IgG específica contra Plasmodium falciparum en sueros de pacientes diagnosticados por gota gruesa con malaria no complicada por P. falciparum, empleando como antígeno un extracto proteico obtenido a partir de cultivo de P. falciparum o un péptido sintético derivado de la proteína de superficie de merozoito GLURP (del inglés: glutamate rich protein). Materiales y métodos. Para la estandarización de la técnica, se utilizaron 22 sueros de pacientes positivos para malaria por P. falciparum y 11 diagnosticados positivos para malaria por P. vivax utilizando la técnica de gota gruesa. Como controles negativos se utilizaron 44 sueros de individuos sanos. Los sueros fueron probados contra extracto de proteínas del parásito y el péptido sintético IMT 94 derivado de la proteína GLURP, para evaluar las concentraciones y las diluciones óptimas de cada componente del sistema. Para la validación de la técnica se utilizaron 251 sueros de pacientes positivos para P. falciparum y 44 sueros de individuos sanos, diagnosticados utilizando la técnica de gota gruesa. Resultados. La técnica estandarizada con el péptido sintético permitió observar diferencia significativa en el reconocimiento de sueros de pacientes, controles positivos y negativos por los antígenos (extracto de proteínas y péptidos sintéticos). Conclusiones. La metodología usada permite identificar la respuestas inmune específica contra P. falciparum.

Abstract Introduction. Malaria continues being one of the diseases causing the greatest morbi-mortality around the world. For that reason, effective diagnostic tools must thus be developed which can be used in strategies for controlling the disease. Objectives. To standardise enzyme- linked immunosorbent assay (ELISA) conditions for detecting Plasmodium falciparum specific IgG in sera from patients diagnosed by thick smear as suffering non-complicated malaria caused by P. falciparum. A protein extract obtained from P. falciparum culture or a synthetic peptide derived from glutamate rich protein (GLURP) merozoite surface protein would be used as antigen. Materials and Methods. 22 serum samples from patients diagnosed as suffering from P. falciparum malaria, 11 serum samples from patients diagnosed as suffering from P. vivax and 44 from healthy donors, diagnosed by using the thick smear tecnique were used for standarising the technique. Serum samples were tested against parasite protein extract and GLURP- derived IMT 94 synthetic peptide for standardisign optimum dilutions and concentrations for each component in the system. 251 serum samples from patients diagnosed as suffering from P. falciparum malaria and 44 from healthy donors diagnosed by using the thick smear tecnique were used to validate the technique. Results. The technique led to significant differences being observed in antigens (protein extract and synthetic peptides) recognising serum from positive and negative patients and controls. Conclusions. The methodology used led to identifying specific immune response against P. falciparum.

A GBP 130 derived peptide from Plasmodium falciparum binds to human erythrocytes and inhibits merozoite invasion in vitro

The malarial GBP 130 protein binds weakly to intact human erythrocytes; the binding sites seem to be located in the repeat region and this region's antibodies block the merozoite invasion. A peptide from this region (residues from 701 to 720) which binds to human erythrocytes was identified. This peptide named 2220 did not bind to sialic acid; the binding site on human erythrocyte was affected by treatment with trypsin but not by chymotrypsin. The peptide was able to inhibit Plasmodium falciparum merozoite invasion of erythrocytes. The residues F701, K703, L705, T706, E713 (FYKILTNTDPNDEVERDNAD) were found to be critical for peptide binding to erythrocytes.