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

Area of Science:

• Veterinary Medicine
• Biochemistry
• Clinical Pathology

Background:

• Hypercoagulation, a state of increased clotting tendency, can occur in horses.
• Thrombin-antithrombin III (TAT) complexes are indicators of coagulation activation.
• Accurate quantification of TAT complexes is crucial for diagnosing hypercoagulable states.

Purpose of the Study:

• To adapt a human enzyme-linked immunosorbent assay (ELISA) kit for quantifying equine thrombin-antithrombin III (TAT) complexes.
• To characterize the adapted assay for reliability and establish reference intervals.
• To evaluate TAT complexes as a potential biomarker for hypercoagulation in horses.

Main Methods:

• A commercially available human TAT ELISA kit was adapted using purified equine thrombin and antithrombin III for standard curves.
• Equine plasma samples from healthy horses (n=29) and ill horses (n=4) were analyzed.
• Assay validation included determination of reference intervals and intra- and inter-assay coefficients of variation.

Main Results:

• The adapted human ELISA kit successfully quantified TAT complexes in equine plasma.
• Reference intervals for TAT concentration in healthy horses were established (mean 3.95 ± 1.93 µg/L).
• Significantly elevated TAT concentrations were observed in horses with conditions associated with hypercoagulation, such as cecal perforation (174.30 µg/L) and endotoxemia (62.98 µg/L).

Conclusions:

• Human TAT ELISA kits are suitable for measuring TAT concentrations in equine plasma.
• TAT complexes serve as a valuable marker for detecting and characterizing hypercoagulation in horses.
• This assay can aid in the clinical management and understanding of equine hypercoagulable states.