description
- Coeliac disease is an inflammatory disease of the upper small intestine that affects as much as 1% of the European population and results from gluten ingestion in genetically susceptible individuals. The only treatment for CD is a strict gluten-free diet, with the longer the individual fails to adhere to this diet, the greater the chance of developing malnutrition and other complications. Thus, the existence of reliable gluten free food is crucial to the well-being of the population. Current assays available for detection of gluten in food suffer from many drawbacks. Firstly, there appears increasing knowledge as to the toxicity of particular gluten proteins. Secondly, there is no existing protocol that has been demonstrated to quantitatively extract gluten from both raw and processed foodstuffs and assays based on the use of antibodies are incompatible with the reducing reagents. Fourthly, assays typically used for gluten detection exploit indirect sandwich assays, typically requiring 6-8 hours to complete. Here we propose to address each of the challenges to develop reagentless biosensors for in-situ detection of gluten in foodstuffs by using aptamers. Aptamers are getting more and more attention in biosensor area especially because of their superior properties over antibodies. Aptamers will be selected against these identified gluten toxic sequences in buffer conditions akin to that required for gluten extraction and are thus unaffected by the reducing agents present (contrary to the case of antibodies). Furthermore, aptamers lend themselves to highly flexible assay formats not attainable using antibodies. In the work proposed here, we intend to take special extraction protocol and selected aptamers and exploit them for use in various reagentless aptasensor formats (e.g. fluorescent-electrochemical displacement, molecular aptamer beacons) that will allow the rapid, inexpensive, accurate and facile detection of gluten in foodstuffs.