2009 Annual Report 1a.Objectives (from AD-416) The objective of this cooperative research project is to develop multiplex peptide nucleotide acid (PNA) chip and immune biomarkers associated with food-borne pathogens. 1b.Approach (from AD-416) NVRQS will develop multiplex PNA chip by identifying SNP of ffod-borne pathogens and validate PNA chip for diagnosis and detection of Clostridium, Salmonella and Campylobacter. ARS will develop disease challenge models to evaluate vaccines against poultry food-poisoning pathogen, develop and optimize real-time RT-PCR for detection of C. septicum and C. perfringens, develop ELISA for chicken cytokine detection to identify immune biomarkers associated with C. perfringens and C. septicum and evaluate potential vaccine candidates of Clostricium proteins. 3.Progress Report The major objectives of this collaboration are: to develop multiplex peptide nucleotide acid (PNA) chip based on single nucleotide polymorphism (SNP) that can detect and diagnose major food-borne pathogen species and practical enzyme-linked immunosorbent assay (ELISA) system to detect major chicken cytokines associated with innate immune responses of poultry to food-poisoning pathogens, and to develop biomarkers associated with host immune responses to major food-borne pathogen species including Clostridium species. The first part is being carried out by NVRQS and ARS is currently identifying immunogenic proteins of Clostridium for ELISA development.