Effect of Pre-analytical Conditions on Prothrombin Time and Partial Activated Thromboplastin Time

Author(s): Moutinho Beatriz , Pinto Beatriz , Cardoso Rita , Alves Helena , Botelho Monica* .

Journal Name: Current Pharmaceutical Biotechnology

Volume 20 , Issue 4 , 2019

Become EABM
Become Reviewer

Graphical Abstract:


Abstract:

Background: Clinical analysis often involves clotting assays. Although the guidelines suggest the storing and freezing of samples before these assays, there are contradictory results in the literature. The objective of this study was to analyse the effect of the temperature and the storage of plasma sample on Prothrombin Time (PT) and Activated Partial Thromboplastin Time (aPTT) in clinical samples for 65 patients without coagulation disorders.

Materials and Methods: After centrifugation, plasma of each patient was tested at arrival as part of their routine care and separately aliquoted. Three aliquots were stored at room temperature, 4°C and - 20°C for 24h after collection, two aliquots were stored at 4°C and -20°C for 1 week and one aliquot was stored at -70°C for 1 month.

Results: PT from healthy patients was affected at room temperature for 24h and at 4°C for 1 week. For aPTT, the results were statistically different for all the conditions after 24h and at 4°C for 1 week.

Conclusion: Results indicate that PT and aPTT can be stored at -70ºC for at least 1 month without any significant changes in the assay result.

Keywords: Coagulation, plasma, PT, aPTT, pre-analytical, sample analysis.

[1]
Mann, K.G.; Lorand, L. Introduction: Blood coagulation. Methods Enzymol., 1993, 222, 1-10.
[2]
O’Connor, S.D.; Taylor, A.J.; Williams, E.C.; Winter, T.C. Coagulation concepts update. Am. J. Roentgenol., 2009, 193, 1656-1664.
[3]
Gosselin, R.C.; Dwyre, D.W. Determining the effect of freezing on coagulation testing: Comparasion of results between fresh and once frozen-thawed plasma. Blood Coagul. Fibrinolysis, 2015, 26, 69-74.
[4]
Alesci, S.; Borggrefe, M.; Dempfle, C.E. Effect of freezing method and storage at -20ºC and -70ºC on prothrombin time, aPTT and plasma fibrinogen levels. Thromb. Res., 2009, 124, 121-126.
[5]
Funk, D.M.A.; Lippi, G.; Favoloro, E.J. Quality standands for sample processing, transportantion, and storage in hemostasis testing. Semin. Thromb. Hemost., 2012, 38, 576-585.
[6]
Favaloro, E.J.; Funk, D.M.; Lippi, G. Pre-analytical variables in coagulation testing associated with diagnostic errors in hemostasis. Labmedicine, 2012, 43, 1-10.
[7]
CLSI. Collection, transport, and processing of blood specimens for testing plasma-based coagulation assays and molecular hemostasis assays; Approved guideline - fifth edition, Wayne, PA: Clinical and Laboratory Standards Institute: CLSI document H21 - A4, 2008.
[8]
Totzke, U.; Kuyas, C. Non-frozen transports of whole blood samples do not cause relevant bias for global coagulation tests in clinical trials evaluating the drug safety. Contemp. Clin. Trials, 2005, 26, 488-502.
[9]
Thiele, T.; Hron, G.; Kellner, S.; Wasner, C.; Westphal, A.; Warkentin, T.; Greinacher, A.; Selleng, K. Thrombin generation, ProC®Global, prothrombin time and activated partial thromboplastin time in thawed plasma stored for seven days and after methylene blue/light pathogen inactivation. Blood Transfus., 2016, 14, 66-72.
[10]
Lamboo, M.; Poland, D.C.W.; Eikenboom, J.C.J.; Harvey, M.S.; Groot, E.; Brand, A.; de Vries, R.R. Coagulation parameters of thawed fresh-frozen plasma during storage at different temperatures. Transfus. Med., 2007, 17, 182-186.
[11]
Lippi, G.; Rossi, R.; Ippolito, L.; Zobbi, V.; Azzi, D.; Pipitone, S.; Favaloro, E.J.; Funk, D.M. Influence of residual platelet count on routine coagulation, factor VIII, and Factor IX testing in postfreeze-thaw samples. Semin. Thromb. Hemost., 2013, 39, 834-839.
[12]
CLSI. One-Stage Prothrombin Time (PT) Test and Activated Partial Thromboplastin Time (APTT) Test; Approved Guideline - Second Edition,, PA: Clinical and Laboratory Standards Institute: CLSI document H47-A2, 2009.
[13]
Foshat, M.; Bates, S.; Russo, W.; Huerta, A.; Albright, K.; Giddings, K.; Indrikovs, A.; Qian, Y.W. Effect of freezing plasma at -20ºC for 2 weeks on prothrombin time, activated partial thromboplastin time, dilute russell viper venom time, activated protein C resistance, and D-Dimer levels. Clin. Appl. Thromb. Hemost., 2015, 21, 41-47.
[14]
Gosselin, R.C.; Honeychurch, K.; Dwyre, D.M. Effects of storage and thawing conditions on coagulation testing. Int. J. Lab. Hematol., 2015, 37, 551-559.


Rights & PermissionsPrintExport Cite as

Article Details

VOLUME: 20
ISSUE: 4
Year: 2019
Page: [327 - 331]
Pages: 5
DOI: 10.2174/1389201020666190314125918
Price: $58

Article Metrics

PDF: 20
HTML: 3