Enzyme-linked immunosorbent assay for thrombin-antithrombin III complexes in horses.

This research aimed to adapt a human enzyme-linked immunosorbent assay (ELISA) kit to analyze thrombin-antithrombin III (TAT) complexes in horses, aiming to establish TAT as an indicator for a hypercoagulable state in horses.

Objective and Methodology

• The study aimed to adapt and characterize a human ELISA kit to quantify TAT complexes specifically in equine blood samples. The idea was to evaluate if TAT could be an indicator of hypercoagulation in horses.
• Two sets of horses were utilized in this research. A control group comprised 29 clinically normal horses, while the second group included 4 ill horses known to have causes related to hypercoagulation.
• An ELISA kit, typically used for humans and carrying two antibodies against human thrombin and antithrombin III, was adapted for the study. These antibodies selectively bind to their corresponding TAT antigenic sites.
• Standards for equine TAT were made from purified equine thrombin and antithrombin III, diluted in a phosphate-buffered saline solution. The solution contained Tween and heparin to establish standard curves.
• Lastly, reference intervals for TAT concentration in equine plasma with added citrate, and intra- and interassay coefficients of variation were determined.

Results and Conclusions

• The research showed mean values with standard deviations, median results, and the range of TAT concentrations.
• The researchers observed an increase of TAT concentration in ill horses. Notably, a horse with a cecal perforation showed a significant increase in TAT concentration, as did another horse that had been infused with endotoxin.
• Consequently, the research data suggested TAT as a firm indicator of hypercoagulation in horses.
• The conclusions drawn from this study imply that human ELISA kits can be effectively adapted to measure TAT concentration in horses and might help further characterize hypercoagulable conditions in equine medical practice.