Development of diagnostic teststrips incorporating concatameric aptamers for quantitative detection of C. difficile

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The emergence of new pathogenic species and strains that are resistant to traditional treatments are amongst the most pressing health concerns of our time. The bird flu virus epidemic and the outbreak of Clostridium difficile in several hospitals across Canada in the past year are the latest examples to drive this point home. These outbreaks can be a huge economic burden and, more importantly, pose a tremendous risk to human health. Early and accurate detection and identification of dangerous pathogens is a crucial step in preventing large-scale outbreaks. This is particularly relevant today as globalization of commerce and decreased travel times have significantly increased the rate and breadth of the spread of infectious agents. Traditional microbial pathogen detection techniques using molecular methods (DNA detection with PCR) or immunological methods require large pathogen concentrations and/or long analysis times - time that can be very precious when assessing a patient with a deteriorating condition or when trying to identify and control a large-scale outbreak. In addition, current methods often cannot distinguish between carriers of a disease and those who are infective (a particular issue with C. difficile). Hence there is still a significant need to develop rapid, sensitive, accurate and inexpensive methods and devices for the detection of pathogens. Equally important is the need for a reliable method that can be put in place quickly to create new sensors and assays to respond to unanticipated outbreaks. The goal of this CRD project is to develop a platform based on nucleic acid aptamers and enzymes that can provide high amplification of a selective binding event between the nucleic acid and the pathogen to allow detection of very low levels of pathogens in minutes on a simple paper-based test strip. The proposal brings together two co-applicants with expertise in DNA aptamer and DNA enzyme technology, biomolecule immobilization and biosensors, and development of bioactive paper-based diagnostics. This team will develop and validate new aptamer and aptazyme sensing platforms for detection of pathogens that can be licensed by our Canadian partner for eventual commercialization.