Development and evaluation of indirect enzyme-linked immunosorbent assays for the determination of immune response to multiple clostridial antigens in vaccinated captive bred southern white rhinoceros (Ceratotherium simum simum)

Table 1 Summary of the iELISA antigens used for assay development

Clostridium species	Source	Antigen
C. perfringens type A	Sigma Aldrich	Phospholipase C^b
C. chauvoei	Aphis, USDA*	Flagella antigen
C. septicum	Aphis, USDA*	Culture filtrate containing ATX used for MNT by USDA^a
C. sordellii	Aphis, USDA*	Culture filtrate containing TcsL used for MNT by USDA^a
C. novyi	NIBSC (COT)^c	Culture filtrate containing TcnA used for MNT by USDA^d

Specific antigens used to coat the plates for the different iELISAs are described as well as their sources
^*WHO International Standard routinely used for ELISA and or mouse neutralization tests (MNT) donated by the USDA
^aWHO International ELISA Standard donated by the United States Department of Agriculture (USDA) received as formalinized inactivated antigens
^bPhospholipase C (P4039, Sigma Aldrich), 125 units per mg lyophilized protein phospholipase C expressed from C. perfringens strain 13, plc(988262), chromatographically purified [20]. Phospholipase C may not be the only antigen of importance, but the decision was taken to at least evaluate one target protein per organism based on recommendations in publications. Phospholipase C evaluation was made based on McCourt et al. [20]
^cInternationally prescribed C. novyi alpha toxin from the NIBSC as prescribed by the European Pharmacopoeia [21]
^dNational Institute for Biological Standards and Control, NIBSC, received as formalinized inactivated antigens

ISSN: 1751-0147