Abstract
Background
Correct handling and storage of blood samples for coagulation tests are important to assure correct diagnosis and monitoring. The aim of this study was to assess the pre-analytical practices for routine coagulation testing in European laboratories.
Methods
In 2013–2014, European laboratories were invited to fill in a questionnaire addressing pre-analytical requirements regarding tube fill volume, citrate concentration, sample stability, centrifugation and storage conditions for routine coagulation testing (activated partial thromboplastin time [APTT], prothrombin time in seconds [PT-sec] and as international normalised ratio [PT-INR] and fibrinogen).
Results
A total of 662 laboratories from 28 different countries responded. The recommended 3.2% (105–109 mmol/L) citrate tubes are used by 74% of the laboratories. Tube fill volumes ≥90% were required by 73%–76% of the laboratories, depending upon the coagulation test and tube size. The variation in centrifugation force and duration was large (median 2500 g [10- and 90-percentiles 1500 and 4000] and 10 min [5 and 15], respectively). Large variations were also seen in the accepted storage time for different tests and sample materials, for example, for citrated blood at room temperature the accepted storage time ranged from 0.5–72 h and 0.5–189 h for PT-INR and fibrinogen, respectively. If the storage time or the tube fill requirements are not fulfilled, 72% and 84% of the respondents, respectively, would reject the samples.
Conclusions
There was a large variation in pre-analytical practices for routine coagulation testing in European laboratories, especially for centrifugation conditions and storage time requirements.
Acknowledgments
The authors thank EQALM and the national EQA organisations for distributing the questionnaire to their members, and all participating laboratories for their time and thoughtful completion of the questionnaire. We also thank Andreas Hillarp for valuable comments on the manuscript and the Norwegian Western Health Authority for post-doc scholarship for Ann Helen Kristoffersen.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organisation(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
References
1. Favaloro EJ, Adcock Funk DM, Lippi G. Pre-analytical variables in coagulation testing associated with diagnostic errors in hemostasis. Lab Med 2012;43:1–10.10.1309/LM749BQETKYPYPVMSearch in Google Scholar
2. Favaloro EJ, Lippi G, Adcock DM. Preanalytical and postanalytical variables: the leading causes of diagnostic error in hemostasis? Semin Thromb Hemost 2008;34:612–34.10.1055/s-0028-1104540Search in Google Scholar PubMed
3. Adcock Funk DM, Lippi G, Favaloro EJ. Quality standards for sample processing, transportation, and storage in hemostasis testing. Semin Thromb Hemost 2012;36:576–85.10.1055/s-0032-1319768Search in Google Scholar PubMed
4. Adcock DM, Kressin DC, Marlar RA. Minimum specimen volume requirements for routine coagulation testing: dependence on citrate concentration. Am J Clin Pathol 1998;109:595–9.10.1093/ajcp/109.5.595Search in Google Scholar PubMed
5. Zurcher M, Sulzer I, Barizzi G, Lammle B, Alberio L. Stability of coagulation assays performed in plasma from citrated whole blood transported at ambient temperature. Thromb Haemost 2008;99:416–26.10.1160/TH07-07-0448Search in Google Scholar PubMed
6. Linskens EA, Devreese KM. Pre-analytical stability of coagulation parameters in plasma stored at room temperature. Int J Lab Hematol 2018;40:292–303.10.1111/ijlh.12784Search in Google Scholar PubMed
7. CLSI. Collection, transport, and processing of blood specimens for testing plasma-based coagulation assays and molecular hemostasis assays, Approved Guideline – Fifth Edition. CLSI document H21-A5. Wayne, PA: Clinical and Laboratory Standards Institute, 2008.Search in Google Scholar
8. Lippi G, Salvagno GL, Montagnana M, Lima-Oliveira G, Guidi GC, Favaloro EJ. Quality standards for sample collection in coagulation testing. Semin Thromb Hemost 2012;38:565–75.10.1055/s-0032-1315961Search in Google Scholar PubMed
9. Polack B, Schved JF, Boneu B, Groupe d’Etude sur l’Hemostase et la T. Preanalytical recommendations of the ‘Groupe d’Etude sur l’Hemostase et la Thrombose’ (GEHT) for venous blood testing in hemostasis laboratories. Haemostasis 2001;31:61–8.10.1159/000048046Search in Google Scholar PubMed
10. Sectie Stolling van de Stichting Kwaliteitsbewaking Medische Laboratoriumdiagnostiek. Preanalytische voorschriften voor stollingsbepalingen, 2012. Available at: https://www.nvkc.nl/sites/default/files/11.pdf. Accessed: 2019.Search in Google Scholar
11. Mackie I, Cooper P, Lawrie A, Kitchen S, Gray E, Laffan M, et al. Guidelines on the laboratory aspects of assays used in haemostasis and thrombosis. Int J Lab Hematol 2013;35:1–13.10.1111/ijlh.12004Search in Google Scholar PubMed
12. Use of anticoagulants in diagnostic laboratory investigations and Stability of blood, plasma and serum samples, 2002. Available at: https://apps.who.int/iris/bitstream/ handle/10665/65957/WHO_DIL_LAB_99.1_REV.2.pdf. Accessed: 2019.Search in Google Scholar
13. Lippi G, Plebani M, Favaloro EJ. Interference in coagulation testing: focus on spurious hemolysis, icterus, and lipemia. Semin Thromb Hemost 2013;39:258–66.10.1055/s-0032-1328972Search in Google Scholar PubMed
14. Magnette A, Chatelain M, Chatelain B, Ten Cate H, Mullier F. Pre-analytical issues in the haemostasis laboratory: guidance for the clinical laboratories. Thromb J 2016;14:49.10.1186/s12959-016-0123-zSearch in Google Scholar PubMed PubMed Central
15. Stavelin A, Albe X, Meijer P, Sarkany E, MacKenzie F. An overview of the European Organization for External Quality Assurance Providers in Laboratory Medicine (EQALM). Biochem Med (Zagreb) 2017;27:30–6.10.11613/BM.2017.005Search in Google Scholar PubMed PubMed Central
16. Lippi G, Salvagno GL, Adcock DM, Gelati M, Guidi GC, Favaloro EJ. Right or wrong sample received for coagulation testing? Tentative algorithms for detection of an incorrect type of sample. Int J Lab Hematol 2010;32:132–8.10.1111/j.1751-553X.2009.01142.xSearch in Google Scholar PubMed
17. Adcock DM, Kressin DC, Marlar RA. Effect of 3.2% vs. 3.8% sodium citrate concentration on routine coagulation testing. Am J Clin Pathol 1997;107:105–10.10.1093/ajcp/107.1.105Search in Google Scholar PubMed
18. Duncan EM, Casey CR, Duncan BM, Lloyd JV. Effect of concentration of trisodium citrate anticoagulant on calculation of the International Normalised Ratio and the International Sensitivity Index of thromboplastin. Thromb Haemost 1994;72:84–8.10.1055/s-0038-1648816Search in Google Scholar
19. Reneke J, Etzell J, Leslie S, Ng VL, Gottfried EL. Prolonged prothrombin time and activated partial thromboplastin time due to underfilled specimen tubes with 109 mmol/L (3.2%) citrate anticoagulant. Am J Clin Pathol 1998;109:754–7.10.1093/ajcp/109.6.754Search in Google Scholar PubMed
20. Pretorius L, Janse van Rensburg WJ, Conradie C, Coetzee MJ. Minimum citrate tube fill volume for routine coagulation testing. Int J Lab Hematol 2014;36:493–5.10.1111/ijlh.12155Search in Google Scholar PubMed
21. Ver Elst K, Vermeiren S, Schouwers S, Callebaut V, Thomson W, Weekx S. Validation of the minimal citrate tube fill volume for routine coagulation tests on ACL TOP 500 CTS(R). Int J Lab Hematol 2013;35:614–9.10.1111/ijlh.12099Search in Google Scholar PubMed
22. Chuang J, Sadler MA, Witt DM. Impact of evacuated collection tube fill volume and mixing on routine coagulation testing using 2.5-mL (pediatric) tubes. Chest 2004;126:1262–6.10.1378/chest.126.4.1262Search in Google Scholar PubMed
23. Lippi G, Salvagno GL, Montagnana M, Manzato F, Guidi GC. Influence of the centrifuge time of primary plasma tubes on routine coagulation testing. Blood Coagul Fibrinolysis 2007;18:525–8.10.1097/MBC.0b013e3281eec945Search in Google Scholar PubMed
24. Sultan A. Five-minute preparation of platelet-poor plasma for routine coagulation testing. East Mediterr Health J 2010;16: 233–6.10.26719/2010.16.2.233Search in Google Scholar
25. Dimeski G, Solano C, Petroff MK, Hynd M. Centrifugation protocols: tests to determine optimal lithium heparin and citrate plasma sample quality. Ann Clin Biochem 2011;48:218–22.10.1258/acb.2010.010230Search in Google Scholar PubMed
26. Nelson S, Pritt A, Marlar RA. Rapid preparation of plasma for ‘Stat’ coagulation testing. Arch Pathol Lab Med 1994;118:175–6.Search in Google Scholar
27. Baglin T, Luddington R. Reliability of delayed INR determination: implications for decentralized anticoagulant care with off-site blood sampling. Br J Haematol 1997;96:431–4.10.1046/j.1365-2141.1997.d01-2079.xSearch in Google Scholar PubMed
28. Heil W, Grunewald R, Amend M, Heins M. Influence of time and temperature on coagulation analytes in stored plasma. Clin Chem Lab Med 1998;36:459–62.10.1515/CCLM.1998.077Search in Google Scholar PubMed
29. Salvagno GL, Lippi G, Montagnana M, Franchini M, Poli G, Guidi GC. Influence of temperature and time before centrifugation of specimens for routine coagulation testing. Int J Lab Hematol 2009;31:462–7.10.1111/j.1751-553X.2008.01058.xSearch in Google Scholar PubMed
30. Palmer RN, Gralnick HR. Cold-induced contact surface activation of the prothrombin time in whole blood. Blood 1982;59:38–42.10.1182/blood.V59.1.38.38Search in Google Scholar
31. Favaloro EJ, Soltani S, McDonald J. Potential laboratory misdiagnosis of hemophilia and von Willebrand disorder owing to cold activation of blood samples for testing. Am J Clin Pathol 2004;122:686–92.10.1309/E4947DG48TVY19C2Search in Google Scholar
32. Bohm M, Taschner S, Kretzschmar E, Gerlach R, Favaloro EJ, Scharrer I. Cold storage of citrated whole blood induces drastic time-dependent losses in factor VIII and von Willebrand factor: potential for misdiagnosis of haemophilia and von Willebrand disease. Blood Coagul Fibrinolysis 2006;17:39–45.10.1097/01.mbc.0000198990.16598.85Search in Google Scholar PubMed
33. Narayanan S. Preanalytical aspects of coagulation testing. Haematologica 1995;80:1–6.Search in Google Scholar
34. Cadamuro J, Simundic AM, Ajzner E, Sandberg S. A pragmatic approach to sample acceptance and rejection. Clin Biochem 2017;50:579–81.10.1016/j.clinbiochem.2017.02.001Search in Google Scholar PubMed
Supplementary Material
The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2019-0214).
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