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 food-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 for this reporting period were to compare genetic line differences in disease response to Clostridium infection and to develop a practical enzyme-linked immunosorbent assay (ELISA) system to detect major chicken cytokines associated with innate immune responses to food-poisoning pathogens including Clostridium species. Several chicken cytokine genes and hybridomas which produce mouse monoclonal antibodies to major cytokines were produced, characterized, and provided to NVRQS for development of an ELISA. To develop the necrotic enteritis disease model, two major commercial broiler chicken lines (designated R and C) were compared with respect to their disease susceptibility to C. perfringens (CP) infection. In general, the R breed tended to produce higher serum titers against CP compared to C strain. Significant differences in CP strain on the host antibody response were observed. The results of in vivo trials will be summarized for publication.