Reproducibility, stability, and biological variability of thrombin generation using calibrated automated thrombography in healthy dogs

Vet Clin Pathol. 2018 Jun;47(2):218-226. doi: 10.1111/vcp.12602. Epub 2018 Mar 26.

Abstract

Background: Thrombin plays a central role in hemostasis and thrombosis. Calibrated automated thrombography (CAT), a thrombin generation assay, may be a useful test for hemostatic disorders in dogs.

Objectives: To describe CAT results in a group of healthy dogs, and assess preanalytical variables and biological variability.

Animals: Forty healthy dogs were enrolled.

Methods: Lag time (Lag), time to peak (ttpeak), peak thrombin generation (peak), and endogenous thrombin potential (ETP) were measured. Direct jugular venipuncture and winged-needle catheter-assisted saphenous venipuncture were used to collect samples from each dog, and results were compared between methods. Sample stability at -80°C was assessed over 12 months in a subset of samples. Biological variability of CAT was assessed via nested ANOVA using samples obtained weekly from a subset of 9 dogs for 4 consecutive weeks.

Results: Samples for CAT were stable at -80°C over 12 months of storage. Samples collected via winged-needle catheter venipuncture showed poor repeatability compared to direct venipuncture samples; there was also poor agreement between the 2 sampling methods. Intra-individual variability of CAT parameters was below 25%; inter-individual variability ranged from 36.9% to 78.5%.

Conclusions: Measurement of thrombin generation using CAT appears to be repeatable in healthy dogs, and samples are stable for at least 12 months when stored at -80°C. Direct venipuncture sampling is recommended for CAT. Low indices of individuality suggest that subject-based reference intervals are more suitable when interpreting CAT results.

Keywords: Canine; hemostasis.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Biological Variation, Population
  • Blood Coagulation Tests / veterinary*
  • Dogs
  • Female
  • Male
  • Prospective Studies
  • Reproducibility of Results
  • Thrombin / metabolism*
  • Thrombosis / veterinary*

Substances

  • Thrombin