Original scientific paper

https://doi.org/10.11613/BM.2015.022

Plasma centrifugation does not influence thrombin-antithrombin and plasmin-antiplasmin levels but determines platelet microparticles count

Ewa Stępień orcid.org/0000-0003-3589-1715 ; Department of Clinical Biochemistry, Jagiellonian University Medical College, Krakow, Poland
Krzysztof Gruszczyński ; Department of Molecular Diagnostics, Holycross Cancer Centre, Kielce, Poland
Przemysław Kapusta ; Laboratory for Molecular Biology and Research, John Paul II Hospital, Krakow, Poland
Artur Kowalik orcid.org/0000-0002-3718-999X ; Department of Molecular Diagnostics, Holycross Cancer Centre, Kielce, Poland
Iwona Wybrańska ; Department of Clinical Biochemistry, Jagiellonian University Medical College, Krakow, Poland

Abstract

Introduction: Centrifugation is an essential step for plasma preparation to remove residual elements in plasma, especially platelets and platelet-derived microparticles (PMPs). Our working hypothesis was that centrifugation as a preanalytical step may influence some coagulation parameters.
Materials and methods: Healthy young men were recruited (N = 17). For centrifugation, two protocols were applied: (A) the first centrifugation at 2500 x g for 15 min and (B) at 2500 x g for 20 min at room temperature with a light brake. In protocol (A), the second centrifugation was carried out at 2500 x g for 15 min, whereas in protocol (B), the second centrifugation involved a 10 min spin at 13,000 x g. Thrombin-antithrombin (TAT) and plasmin-antiplasmin (PAP) complexes concentrations were determined by enzyme-linked immunosorbent assays. PMPs were stained with CD41 antibody and annexin V, and analyzed by flow cytometry method. Procoagulant activity was assayed by the Calibrated Automated Thrombogram method as a slope of thrombin formation (CAT velocity).
Results: Median TAT and PAP concentrations did not differ between the centrifugation protocols. The high speed centrifugation reduced the median (IQR) PMP count in plasma from 1291 (841-1975) to 573 (391-1010) PMP/µL (P = 0.001), and CAT velocity from 2.01 (1.31-2.88) to 0.97 (0.82-1.73) nM/min (P = 0.049). Spearman’s rank correlation analysis showed correlation between TAT and PMPs in the protocol A plasma which was (rho = 0.52, P < 0.050) and between PMPs and CAT for protocol A (rho = 0.74, P < 0.050) and protocol B (rho = 0.78, P < 0.050).
Conclusion: Centrifugation protocols do not influence the markers of plasminogen (PAP) and thrombin (TAT) generation but they do affect the PMP count and procoagulant activity

Keywords

cell-derived microparticles; blood coagulation tests; centrifugation; preanalytical phase

Hrčak ID:

139807

URI

https://hrcak.srce.hr/139807

Publication date:

15.6.2015.

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