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. 2022 Jun;44(3):610-618.
doi: 10.1111/ijlh.13794. Epub 2022 Jan 13.

Stability of coagulation parameters in plasma samples at room temperature after one freeze/thaw cycle

Marion Gaudard  1 , Elodie Boissier  2 , Laurie Talon  1 , Jonathan Douxfils  3   4 , Anne-Françoise Sapin  1 , Thomas Sinegre  1   5 , Aurélien Lebreton  1   5
Affiliations

Stability of coagulation parameters in plasma samples at room temperature after one freeze/thaw cycle

Marion Gaudard et al. Int J Lab Hematol. 2022 Jun.

Abstract

Introduction: Sample freezing is a part of routine laboratory tasks because some coagulation parameters are analysed in batches to optimize reagent consumption. The coagulation parameter stability in fresh and frozen samples has been described, but data are scarcer after thawing. This study objective was to determine the stability of the main coagulation parameters (from blood withdrawn on siliconized CTAD tubes and double-centrifuged) after one freeze/thaw cycle to generate procedures for appropriate handling, storage and testing.

Methods: Prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen, D-dimers, clotting factors (F), protein C, protein S, antithrombin, lupus anticoagulant (LA)-sensitive aPTT and diluted-Russel's viper venom time (dRVVT) were assessed in 60 plasma samples (n=30, normal range and n=30, outside the normal range). Thirty samples from anticoagulated patients [unfractionated heparin (UFH), low-molecular weight heparin (LMWH), apixaban or rivaroxaban] were assessed using specific anticoagulant assays. Frozen samples were thawed, and assays were performed at 15 min, 2, 4 and 6 h after thawing. The coagulation parameter stability was assessed with the method of rejection limit.

Results: After thawing, aPTT, PT, fibrinogen, D-dimers, FII, FV, FX, FIX, FXI, FXII, PC and UFH anti-Xa activity remained stable for at least 6 h, FVII for 5 h, PS, AT, dRVVT screen assay and LMWH anti-Xa activity for 4 h, and LA-sensitive aPTT and apixaban-specific anti-Xa activity for 3 h. FVIII, dRVVT confirm assay and rivaroxaban specific anti-Xa activity were stable for 2 h.

Conclusion: These results suggest that sample stability for some haemostasis assays is limited after thawing.

Keywords: anticoagulants; blood coagulation factors; freezing; laboratories; plasma.

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References

REFERENCES

    1. van Geest-Daalderop JHH, Mulder AB, Boonman-de Winter LJM, Hoekstra MMCL, van den Besselaar AMHP. Preanalytical variables and off-site blood collection: influences on the results of the prothrombin time/international normalized ratio test and implications for monitoring of oral anticoagulant therapy. Clin Chem. 2005;51(3):561-568. doi:10.1373/clinchem.2004.043174
    1. Oddoze C, Lombard E, Portugal H. Stability study of 81 analytes in human whole blood, in serum and in plasma. Clin Biochem. 2012;45(6):464-469. doi:10.1016/j.clinbiochem.2012.01.012
    1. Rimac V, Coen HD. Is it acceptable to use coagulation plasma samples stored at room temperature and 4°C for 24 hours for additional prothrombin time, activated partial thromboplastin time, fibrinogen, antithrombin, and D-dimer testing? Int J Lab Hematol. 2017;39(5):475-481. doi:10.1111/ijlh.12664
    1. Heil W, Grunewald R, Amend M, Heins M. Influence of time and temperature on coagulation analytes in stored plasma. Clin Chem Lab Med. 1998;36(7): doi:10.1515/CCLM.1998.077
    1. Toulon P, Metge S, Hangard M, Zwahlen S, Piaulenne S, Besson V. Impact of different storage times at room temperature of unspun citrated blood samples on routine coagulation tests results. Results of a bicenter study and review of the literature. Int J Lab Hematol. 2017;39(5):458-468. doi:10.1111/ijlh.12660

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