Produção de anticorpos para detecção de Escherichia coli enteropatogênica

Abstract

Escherichia coli (EPEC) was frequent cause of many cases of diarrhea in Brazil and around the world, and one of the main agents responsible for high mortality rates among children aged 0-5 years. EPEC is known to cause histological damage to the microvilli of enterocytes, characterized by the formation of a shaped support ultrastructure. One of the factors responsible for pathogenicity is complex BFP (bundle forming pili). The presence of genes BfpA promotes adhesion and consequently the injury of EPEC strains characterized as typical. Differently, some EPEC strains lack EAF (EPECBfpA-) causing lesions shaped as pedestal, but more slowly, called atypical EPEC. BfpA is known as immunogenic and recently the incidence of atypical (EPECBfpA-) is increasing, it is believed that these bacteria injury occurs through other means making it important to develop strategies for identifying. In this study several predicted peptide sequences were synthesized, to produce polyclonal antibodies and anti-EPEC -BfpA in BALB / c antibodies that are used to target specific identification of typical and atypical EPEC. To achieve the objectives of bioinformatics tools and prediction of protein epitopes immuneepitope B were used for antigenic regions specific mapping for the EPEC two phenotypes. Synthetic peptides were purchased to immunize Balb / C mice, so the antisera are evaluated for specificity EPEC. As results, it was possible to construct and synthetize six linear BfpA peptides (P1-P6). P6 peptide was selected as the product of this study due to its ability to induce the antibodies production against this protein and this serum recognize the native protein in typical EPEC strains. We believe that the functional characterization of these antibodies by developing new tools will help in the diagnosis of these two specific strains of EPEC and will contribute to studies involving these bacteria.