Automated synthetic substrate assays for coagulopathies of dogs

Automated synthetic substrate assays for factors II and VII in the extrinsic coagulation pathway have been developed in our laboratory. In both kinetic assays, p-nitroaniline (pNA) was cleaved from the substrate and measured spectrophotometrically at 405 nm. Plasma samples from 5-8 month old beagle dogs were analyzed for factors II and VII, and also for prothrombin times. A subpopulation considered abnormal (prothrombin times greater than 8 seconds) had slightly elevated factor II levels, but extremely low factor VII levels, relative to the normal group. Therefore, the prolonged prothrombin times of this abnormal group can now be confidently attributed to their decreased factor VII levels. The known genetic predisposition of beagle dogs to factor VII deficiency supports this conclusion. Thus, synthetic substrates are useful for the measurement of coagulation factors in dogs, and permit ready automation of the assay.