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Effect of tube filling on plasma freezing for coagulation testing.

Author(s): Sinegre T, Talon L, Sapin AF, Lebreton A

Publication: Res Pract Thromb Haemost, 2023, Vol. 7, Page 100092

PubMed ID: 37065844 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study compared PT; DRVTT; and APTT; von Willebrand factor activity; and levels of Protein C, Protein S, antithrombin, and factors FII, FV, FVII, FVIII, FIX, FX, FXI, and FXII in plasma stored frozen as 0.4, 0.8 and 1.6 mL aliquots in 2 mL tubes. The authors also compared results in plasma stored as 20% filled 2 and 5 mL tubes (0.4 mL and 1 mL, respectively). Blood from 85 patients (diagnosis was not specified, but 7 were receiving unfractionated heparin and 13 vitamin K antagonists) was collected via venipuncture into tubes containing 0.109 M citrate. Platelet poor plasma was separated by centrifugation twice at 2500 g for 15 minutes and stored as 0.4, 0.8 and 1.6 mL aliquots in 2 mL microtubes and as 1 mL aliquots in 5 mL tubes for 3 months at -80°C. Plasma was thawed in a 37°C water bath for 5 min before coagulation testing on a STAR Max coagulometer and AcuStar instrument (von Willebrand factor activity only). 

   Summary of Findings:

   Results of coagulation testing were comparable when plasma was stored as 0.8 mL and 1.6 mL aliquots; however, significant differences were observed when plasma was stored as 0.4 mL aliquots. Compared to the 1.6 mL and 0.8 mL aliquots, 0.4 mL aliquots had shorter PT (P<0.001, both) and aPTT (P<0.001, both); higher levels of fibrinogen (P<0.001, both), FV (P<0.01, both), and antithrombin (P<0.001, and P<0.01, respectively); and lower levels of FVII (P<0.001, both) and FX (P<0.01, both). In addition, the 0.4 mL aliquot had higher levels of FII (P<0.01) than the 1.6 mL aliquot and a longer DRVTT in the screening but not the confirmatory test than the 0.8 mL aliquot (P<0.01). While not significant, the authors report a trend toward an increase in the intrinsic pathway factors (FVIII, FIX and FXII), von Willebrand factor antigen and ristocetin cofactor in the 0.4 mL aliquots relative to the 1.6 mL aliquots, but no change in Protein C or S activity occurred.  In comparison to 0.4 mL aliquots in 2 mL tubes, 1 mL aliquots in 5 mL tubes had a longer aPTT (P<0.001); decreased fibrinogen (P<0.001), FVII (P<0.01), FX (P<0.001), and antithrombin (P<0.01) levels; and higher Protein C activity (P<0.05) but comparable PT, FVIII, FIX, FXI, FXII, Protein S activity and DRVVT screening and confirmatory results.

   Biospecimens

   • Bodily Fluid - Plasma

   Preservative Types

   • Frozen

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   Morphology	Hematology/ auto analyzer
   Protein	Hematology/ auto analyzer
   Glycoprotein	Hematology/ auto analyzer

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage conditions	0.4 mL in 2 mL tube 0.8 mL in 2 mL tube 1.6 mL in 2 mL tube 1.0 mL in 5 mL tube
   Biospecimen Aliquots and Components	Aliquot size/volume	0.4 mL in 2 mL tube 0.8 mL in 2 mL tube 1.6 mL in 2 mL tube 1.0 mL in 5 mL tube

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