Abstract
Interest has recently been renewed in the possible use of Y. 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 of Y. pestis in 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 of Y. pestis from 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 of Y. pestis in food without the need for enrichment. The combined use of the iCropTheBug system and pyrosequencing for efficient capture and detection of Y. pestis is novel and has potential applications in food biodefence.
Highlights
Plague, caused by Yersinia pestis, has given rise to three major pandemics and is considered one of the most devastating diseases in human history [1]
Interest has been renewed in the possible use of Y. pestis as a biological weapon by terrorists, as it could cause mass casualties if dispersed as an aerosol [4]
The vulnerability of food has been demonstrated by the intentional contamination of salad bars in the United States with Salmonella typhimurium, and this makes the possible use of more deadly agents such as Y. pestis a possibility [14]
Summary
Plague, caused by Yersinia pestis, has given rise to three major pandemics and is considered one of the most devastating diseases in human history [1]. The vulnerability of food has been demonstrated by the intentional contamination of salad bars in the United States with Salmonella typhimurium, and this makes the possible use of more deadly agents such as Y. pestis a possibility [14] This concern is exacerbated by the report of multidrug resistant strains [15] and their potential use for bioterrorism in the human population. We present the application of IMS and pyrosequencing based assays for the rapid, specific, and sensitive detection and identification of Y. pestis from food matrices such as milk, bagged salad, and processed meat This assay for Y. pestis detection is a significant improvement over our previous work using the Pathatrix sample preparation system and real-time PCR [22] and demonstrates better limits of detection without an enrichment step. The combination of efficient immunomagnetic concentration of biothreat agents and pyrosequence-based detection system is novel and represents the first report for detection and identification of Y. pestis in food with potential biodefence application
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