2012 Annual Report 1a.Objectives (from AD-416): To develop DNA-based typing/characterization platforms for Shiga toxin-producing E. coli. 1b.Approach (from AD-416): Sequencing of the Escherichia coli O antigen gene clusters and development of genetic-based typing and detection systems. It will address the 60-month milestone for this project, Complete studies on development of genetic-based systems for typing E. coli and detection of non-O157 STEC, and develop methods for isolation of non-O157 STEC. This agreement will expand on our current research and will result in the development of methods to identify, detect, and characterize E. coli O157:H7 and non-O157 STEC, a critical need of regulatory agencies and the food industry. 3.Progress Report: Through a three-way non-funded cooperative agreement, researchers at the ARS ERRC Molecular Characterization of Foodborne Pathogens Research Unit in Wyndmoor, PA collaborated with scientists at the University of Maryland and Life Technologies on research involving DNA sequencing of specific regions in the genome of E. coli strains, including Shiga toxin-producing E. coli (STEC). The sequence data and publicly available genomic information are being utilized to develop rapid genetic-based based detection and identification systems for E. coli and for STEC strains. An ARS scientist is mentoring a graduate student at the University of Maryland who is working on this project. The detection and identification methods are being developed with the cooperation of scientists at Life Technologies into test kits that will benefit the food industry, regulatory agencies, and diagnostic laboratories. The test systems are being evaluated by ARS scientists.