GENERATION OF COLLOIDAL BIOSENSORS FOR THE DETECTION OF Helicobacter pylori IN PATIENT SAMPLES
Resumen
H. pylori is a major cause of gastritis, gastric and duodenal ulcer disease. The genotypes of H. pylori cagA and vacA positive are associated with an increased risk of ulcer and gastric cancer. The project proposes the development of complementary colloidal biosensors with magnetic properties. The first seeks to contribute to the detection of H. pylori bacteria in stool samples, and the second will consist of the detection of antibodies against CagA and VacA in serological samples of patients infected with H.pylori. The procedure consists of searching for conserved amino acid sequences for CagA and VacA in the proteomes of strains isolated in the American continent and the search for the coding sequences for each protein. Next, the conserved coding sequence will be cloned into an expression vector for each protein. The recombinant proteins generated will be expressed and purified. In addition, magnetic nanoparticles (MNP) will be synthesized, and the recombinant proteins (CagA and VacA) or the polyclonal antibodies generated against H. pylori will be immobilized onto the MNP. Finally, the biosensors will be tested with clinical samples from patients infected and not infected, and ELISA and PCR techniques will validate the results.