Analytical and between-subject variation of thrombin generation measured by calibrated automated thrombography on plasma samples

Anne F Kristensen, Søren R Kristensen, Ursula Falkmer, Anna-Marie B Münster, Shona Pedersen

Research output: Contribution to journalJournal articleResearchpeer-review


BACKGROUND: The Calibrated Automated Thrombography (CAT) is an in vitro thrombin generation (TG) assay that holds promise as a valuable tool within clinical diagnostics. However, the technique has a considerable analytical variation, and we therefore, investigated the analytical and between-subject variation of CAT systematically. Moreover, we assess the application of an internal standard for normalization to diminish variation.

METHODS: 20 healthy volunteers donated one blood sample which was subsequently centrifuged, aliquoted and stored at -80 °C prior to analysis. The analytical variation was determined on eight runs, where plasma from the same seven volunteers was processed in triplicates, and for the between-subject variation, TG analysis was performed on plasma from all 20 volunteers. The trigger reagents used for the TG assays included both PPP reagent containing 5 pM tissue factor (TF) and PPPlow with 1 pM TF. Plasma, drawn from a single donor, was applied to all plates as an internal standard for each TG analysis, which subsequently was used for normalization.

RESULTS: The total analytical variation for TG analysis performed with PPPlow reagent is 3-14% and 9-13% for PPP reagent. This variation can be minimally reduced by using an internal standard but mainly for ETP (endogenous thrombin potential). The between-subject variation is higher when using PPPlow than PPP and this variation is considerable higher than the analytical variation.

CONCLUSION: TG has a rather high inherent analytical variation but considerable lower than the between-subject variation when using PPPlow as reagent.

Original languageEnglish
JournalScandinavian Journal of Clinical & Laboratory Investigation
Issue number3
Pages (from-to)175-179
Publication statusPublished - 3. Apr 2018
Externally publishedYes


  • Analytical variation
  • between-subject variation
  • blood coagulation
  • blood coagulation test
  • thrombin
  • thrombin generation


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