Team:Aberdeen Scotland/Project

From 2014.igem.org

Team:Aberdeen Scotland/Project - 2014.ogem.org



E. coli-based Trypanosomiasis Diagnostic System

Getting back at the Sleeping Sickness by detecting it early


E.coli display of peptide mimotopes, integrated with quorum-sensing AND logic, for diagnosis of tropical diseases Trypanosoma.b.gambiense, the causative agent of Trypanosomiasis (African Sleeping Sickness) infects 30,000 people worldwide, with 60 million at disease risk. Unrecognised, this disease is fatal, but accurate diagnosis, relying on detection of patient serum antibodies against two different trypanosomal antigens, allows successful treatment. We engineered autotransporters Antigen 43 and ice-nucleation protein to successfully express surface epitopes in E.coli that mimic two different trypanosomal antigens. Two E.coli strains were generated, each expressing a distinct surface epitope and either a quorum sensing (QS) sender or receiver module respectively. Immune pull-down of each strain, and their subsequent co-culture, produced a quorum signal, indicating successful detection of two distinct antibody populations in a serum sample. We constructed a cheap, Raspberry Pi computer-controlled fluorimeter suitable for developing countries, showing it could detect QS fluorescence output. This novel application of antigen display, and quorum sensing to implement AND logic sensing, facilitates cheap, simple diagnosis of tropical diseases.