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: 1) to develop multiplex peptide nucleotide acid (PNA) chip based on single nucleotide polymorphism (SNP) that can detect and diagnose major food-borne pathogen species; 2) to develop enzyme-linked immunosorbent assays (ELISA) to detect major chicken cytokines associated with innate immune responses of poultry to food-poisoning pathogens; and 3) to develop biomarkers associated with host immune responses to major food-borne pathogen species including Clostridium species. In this report, genes encoding two major C. perfringens virulence factors, alpha-toxin and the NetB (necrotic enteritis B-like) toxin, both of which are implicated in the pathogenesis of necrotic enteritis (NE), were cloned and recombinant protein expressed. Antibodies which detect these two major biomarkers of Clostridium perfringens are being developed and sensitive ELISA assays will be developed to monitor these two toxin proteins during C. perfringens infections. However, their utility as poultry vaccines against Clostridium field infections remains to be realized. Since Clostridium-related poultry diseases such as Gangrenous dermatitis and NE cause substantial economic losses on a global scale, potential vaccine efficacy of these toxin proteins need to be evaluated. This collaboration was monitored by regular meetings and progress reports.