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Development of an Aptamer-Based System for Rapid Identification of Human Noroviruses: Application in Raw Meat Production


<OL> <LI> Develop a laboratory-based aptamer method for detection of human and mouse norovirus capsids. Aptamers for the human and mouse norovirus capsid will be selected from a random DNA pool using Toggle SELEX. Initial selection conditions will be performed in a buffered system with the final selections being performed in filtrates from samples of fecal matter, red meat and juices and retail ground meat.

<LI>Validate an aptamer-based assay (APIK) for use in three experimentally-contaminated matrices. A 'sandwich' method will be developed in 96 well plates. A pool of highly sensitive aptamers will act as the capture aptamers while the most sensitive aptamer will act as the detector aptamer. Initial tests will use high levels of norovirus capsids (i.e. 105 per gram) while subsequent tests will use lower concentrations of capsids to ascertain the detection limit.

<LI>Modify APIK to accommodate spiked unprocessed field samples and test for infectious human and mouse noroviruses. Unprocessed samples will be spiked with infectious human and mouse noroviruses for detection by a modified APIK to accommodate 25 g of sample. Positive tests will be confirmed by plaque assay for mouse norovirus and reverse-transcriptase polymerase chain reaction for human noroviruses. Sensitivity and specificity will be assessed.

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Expected Benefits:
The development of an innovative, rapid norovirus detection method that could be used in assessing the microbiological quality of red meat products throughout the production chain. Such a test could also be readily adapted to detecting norovirus in other meat as well as environmental samples. This test will provide the tool necessary to increase surveillance and thereby the chances of preventing the release of contaminated meat products to the consumer. It will provide the basis for future development of a portable test kit(s) and will demonstrate that aptamer technology can play a role in food surveillance.<P> For more information, please visit the <a href="; target="_blank">Ontario Ministry of Agriculture, Food & Rural Affairs (OMAFRA) Food Safety Research Program</a>.

Sattar , Syed
University of Ottawa
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