 |
 |
 |
 |
 |
 |
29th Annual Meeting and Symposium of the
Desert Tortoise Council, February 20-23, 2004
Abstracts
Chelonian Serodiagnostics: Development of a Field Portable Assay For Detection of Exposure of Tortoises to Mycoplasma agassizii
Daniel R. Brown1, Marianne F. Kramer2, Laurie A.
Zacher1, April M. Green3, and Paul A. Klein3
1College of Veterinary Medicine, University of Florida, Gainesville FL
2Department of Biology, University of South Florida, Tampa FL
3College of Medicine, University of Florida, Gainesville FL
Infectious disease has affected plans for management and conservation of protected chelonians in the United States. Tortoise conservation and recovery plans now include testing for mycoplasmal Upper Respiratory Tract Disease (URTD). Detection of specific anti-mycoplasma antibodies may be used to diagnose infection and immune status of chelonians as a tool for disease management. We have evaluated the feasibility of a field test for specific antibodies against mycoplasma in chelonian plasma, which would provide nearly instant information for management decision making. Preliminary trials were conducted of evanescent-wave biosensor technology for detection of specific anti-Mycoplasma agassizii antibodies in plasma from Gopherus agassizii tortoises. The evanescent-wave biosensor is a laser-based polystyrene fiber optic sensor which detects specific G. agassizii anti-M. agassizii antibody bound to M. agassizii whole-cell lysate antigen. The reporter molecule was Cy5-labeled HL637 monoclonal antibody against tortoise immunoglobulin. Under various experimental protocols, the signals from positive control plasma samples from our bank were three to seven times higher than the signals from negative control plasma samples. A randomized double-blind study was then conducted to determine the sensitivity, specificity, positive predictive value, and negative predictive value of the technique. Preliminary analyses of the results indicate a greater than 90% concordance with the traditional ELISA sample categorization, with a 5 minute per sample, field-portable protocol. Those results suggest that this technology is feasible for application under field conditions. Understanding the dynamics of disease spread in natural wildlife populations may also provide valuable new insights into host:pathogen:population interactions in this era of emerging infectious diseases.
|
|