TY - JOUR
T1 - Analytical and between-subject variation of thrombin generation measured by calibrated automated thrombography on plasma samples
AU - Kristensen, Anne F.
AU - Kristensen, Søren R.
AU - Falkmer, Ursula
AU - Münster, Anna-Marie B.
AU - Pedersen, Shona
PY - 2018
Y1 - 2018
N2 - 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.
AB - 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.
KW - Journal Article
KW - between-subject variation
KW - thrombin
KW - blood coagulation
KW - blood coagulation test
KW - thrombin generation
KW - Analytical variation
KW - Reproducibility of Results
KW - Humans
KW - Middle Aged
KW - Thrombin/analysis
KW - Male
KW - Calibration
KW - Healthy Volunteers
KW - Thrombelastography/methods
KW - Analysis of Variance
KW - Blood Coagulation Tests
KW - Adult
KW - Female
KW - Thromboplastin/chemistry
KW - Observer Variation
UR - http://www.scopus.com/inward/record.url?scp=85041107643&partnerID=8YFLogxK
U2 - 10.1080/00365513.2018.1427280
DO - 10.1080/00365513.2018.1427280
M3 - Journal article
C2 - 29336181
SN - 0036-5513
VL - 78
SP - 175
EP - 179
JO - Scandinavian Journal of Clinical & Laboratory Investigation
JF - Scandinavian Journal of Clinical & Laboratory Investigation
IS - 3
ER -