SUMMARY Necrotic enteritis (NE) is a devastating enteric disease caused by Clostridium perfringens type A/G which impacts global poultry industry by compromising performance, health and welfare of the chickens. The causative main virulent factor responsible for NE pathogenesis has been shifted from a phospholipase C portion of α-toxin, to NE B-like (NetB) toxin, a plasmid-encoded pore-forming heptameric protein, in NE development. Therefore, ability to detect NetB toxin will enable early diagnosis of field NE. The fact that NetB protein can only be detected by western blot analysis using polyclonal anti-NetB antiserum prompted us to develop NetB-specific monoclonal antibody (mAb)-based capture enzyme-linked immunosorbent assay (ELISA). Twenty mAbs reacting with E. coli -expressed NetB protein were selected, isotyped, and conjugated with horseradish peroxidase for antibody pair test. Multiple mAb pairs were found to detect E. coli NetB protein and native NetB protein secreted by netB -positive C. perfringens isolates. The developed capture ELISA could be useful to identify in vitro production of native NetB protein secreted from netB -positive field C. perfringens isolates and to conduct a large field testing of commercial chickens undergoing NE infection. Here, we first report that native NetB toxin can be detected in C. perfringens NetB-specific monoclonal antibody-based capture ELISA.

This content is only available as a PDF.
You do not currently have access to this content.