Rapid Detection and Identification ofYersinia pestisfrom Food Using Immunomagnetic Separation and Pyrosequencing

Abstract

Interest has recently been renewed in the possible use ofY. pestis, the causative agent of plague, as a biological weapon by terrorists. The vulnerability of food to intentional contamination coupled with reports of humans having acquired plague through eating infected animals that were not adequately cooked or handling of meat from infected animals makes the possible use ofY. pestisin a foodborne bioterrorism attack a reality. Rapid, efficient food sample preparation and detection systems that will help overcome the problem associated with the complexity of the different matrices and also remove any ambiguity in results will enable rapid informed decisions to be made regarding contamination of food with biothreat agents. We have developed a rapid detection assay that combines the use of immunomagnetic separation and pyrosequencing in generating results for the unambiguous identification ofY. pestisfrom milk (0.9 CFU/mL), bagged salad (1.6 CFU/g), and processed meat (10 CFU/g). The low detection limits demonstrated in this assay provide a novel tool for the rapid detection and confirmation ofY. pestisin food without the need for enrichment. The combined use of theiCropTheBug system and pyrosequencing for efficient capture and detection ofY. pestisis novel and has potential applications in food biodefence.