PCR-BASED DIAGNOSIS OF HELICOBACTER PYLORI

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Helicobacter pylori (H. pylori) infection is the most frequent bacterial infection worldwide. H. pylori is a major etiological agent in the development of gastritis, gastric ulcer, and gastric cancer. To develop an ultrasensitive, non-invasive screening method for the presence of H. pylori in saliva and other clinical samples, a PCR coamplification assay of urease and beta-globin, followed by fluorometric detection of modified products, is proposed. If the urease gene of H. pylori is present, urease-specific biotin and HEX primers will be physically associated in the double stranded PCR product. Incubation of PCR products in streptavidin coated plates will allow binding of biotinylated primers to streptavidin. The presence of the H. pylori specific PCR product will be evaluated by the detection of fluorescence of the HEX primers. If the screening assay is successful in detecting extremely low levels of H. pylori in the clinical samples, it could be used as a biomarker for early indication of gastric diseases. The commercial application of a screening kit for H. pylori would be tremendous. Proposed commercial applications: This research project for the development of a diagnostic test for the presence of H. pylori could be more sensitive and specific than current methods. The commercial application is tremendous since this is the most frequent bacterial infection worldwide. The methodology is amenable to large scale screening.