Semin Thromb Hemost 2013; 39(03): 258-266
DOI: 10.1055/s-0032-1328972
Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Interference in Coagulation Testing: Focus on Spurious Hemolysis, Icterus, and Lipemia

Giuseppe Lippi
1   Dipartimento di Patologia e Medicina di Laboratorio, U.O. Diagnostica Ematochimica, Azienda Ospedaliero-Universitaria di Parma, Parma, Italy
,
Mario Plebani
2   Dipartimento di Medicina di Laboratorio, Azienda Ospedaliera-Università di Padova, Padova, Italy
,
Emmanuel J. Favaloro
3   Department of Haematology, Institute of Clinical Pathology and Medical Research (ICPMR), Westmead Hospital, Westmead, Australia
› Author Affiliations
Further Information

Publication History

Publication Date:
10 December 2012 (online)

Abstract

The chance that errors might jeopardize the quality of testing is inherently present throughout the total testing process, especially in the preanalytical phase. In the coagulation laboratory, as well as in other areas of diagnostic testing, spurious hemolysis, icteria, and lipemia in test samples represent by far the leading diagnostic challenges. Interference in hemostasis testing due to spurious hemolysis is attributed to both analytical and biologic elements, namely high absorbance of cell-free hemoglobin at wavelengths used by optical instrumentation and release of both cytoplasmatic and plasma membrane molecules (e.g., tissue factor, proteases, phospholipids, and ADP) that can spuriously activate blood coagulation and platelets. The interference attributable to hyperbilirubinemia is mostly due to spectral overlap, whereas that of hypertriglyceridemia mainly reflects elements of light scatter and volume displacement as well as direct interference of lipid particles with hemostasis. In practical terms, spurious hemolysis reflects a more generalized process of endothelial and blood cell damage, so that test results on spuriously hemolyzed specimens should be systematically suppressed. The bias attributable to hyperbilirubinemia is less significant using modern coagulometers equipped with dedicated wavelengths (i.e., with readings at 650 nm or above), so that test results in samples with a bilirubin concentration up to 20 mg/dL can still be analytically reliable. The interference observed in lipemic samples is most evident with readings using wavelengths lower than 500 nm and can hence be prevented with readings at 650 nm or above, and/or using higher dilutions of the test sample, or can be abated in high hypertriglyceridemic specimens (i.e., > 1,000 mg/dL) using high speed microcentrifugation or lipid extraction with organic solvents such as fluorine-chlorinated hydrocarbon, or lipid-clearing agents such as LipoClear (StatSpin Inc., Norwood, MA) and n-hexane.

 
  • References

  • 1 Plebani M, Lippi G. Closing the brain-to-brain loop in laboratory testing. Clin Chem Lab Med 2011; 49 (7) 1131-1133
  • 2 Plebani M, Favaloro EJ, Lippi G. Patient safety and quality in laboratory and hemostasis testing: a renewed loop?. Semin Thromb Hemost 2012; 38 (6) 553-558
  • 3 Lippi G. Governance of preanalytical variability: travelling the right path to the bright side of the moon?. Clin Chim Acta 2009; 404 (1) 32-36
  • 4 Lippi G, Plebani M, Simundic AM. Quality in laboratory diagnostics: from theory to practice. Biochem Med (Zagreb) 2010; 20 (2) 126-130
  • 5 Lippi G, Simundic AM, Mattiuzzi C. Overview on patient safety in healthcare and laboratory diagnostics. Biochem Med (Zagreb) 2010; 20 (2) 131-143
  • 6 Lippi G, Guidi GC, Mattiuzzi C, Plebani M. Preanalytical variability: the dark side of the moon in laboratory testing. Clin Chem Lab Med 2006; 44 (4) 358-365
  • 7 Lippi G, Chance JJ, Church S , et al. Preanalytical quality improvement: from dream to reality. Clin Chem Lab Med 2011; 49 (7) 1113-1126
  • 8 Lippi G, Simundic AM. Total quality in laboratory diagnostics. It's time to think outside the box. Biochem Med (Zagreb) 2010; 20 (1) 5-8
  • 9 Simundic AM, Lippi G. Preanalytical phase—a continuous challenge for laboratory professionals. Biochem Med (Zagreb) 2012; 22 (2) 145-149
  • 10 Salvagno GL, Lippi G, Bassi A, Poli G, Guidi GC. Prevalence and type of pre-analytical problems for inpatients samples in coagulation laboratory. J Eval Clin Pract 2008; 14 (2) 351-353
  • 11 Salvagno GL, Lippi G, Gelati M, Guidi GC. Hemolysis, lipaemia and icterus in specimens for arterial blood gas analysis. Clin Biochem 2012; 45 (4-5) 372-373
  • 12 Favaloro EJ, Adcock Funk DM, Lippi G. Pre-analytical variables in coagulation testing associated with diagnostic errors in hemostasis. Lab Medicine 2012; 43 (2) 1-10
  • 13 Adcock DM, Hoefner DM, Kottke-Marchant K, Marlar RA, Szamosi DI, Warunek DJ. Collection, Transport, and Processing of Blood Specimens for Testing Plasma-Based Coagulation Assays and Molecular Hemostasis Assays; Approved Guideline—Fifth Edition. Wayne, PA: Clinical Laboratory Standards Institute; 2008. . CLSI document H21–A5
  • 14 Lippi G, Blanckaert N, Bonini P , et al. Haemolysis: an overview of the leading cause of unsuitable specimens in clinical laboratories. Clin Chem Lab Med 2008; 46 (6) 764-772
  • 15 Simundic AM, Topic E, Nikolac N, Lippi G. Hemolysis detection and management of hemolysed specimens. Biochem Med (Zagreb) 2010; 20 (2) 154-159
  • 16 Lippi G, Salvagno GL, Montagnana M, Brocco G, Guidi GC. Influence of hemolysis on routine clinical chemistry testing. Clin Chem Lab Med 2006; 44 (3) 311-316
  • 17 Lippi G, Avanzini P, Dipalo M, Aloe R, Cervellin G. Influence of hemolysis on troponin testing: studies on Beckman Coulter UniCel Dxl 800 Accu-TnI and overview of the literature. Clin Chem Lab Med 2011; 49 (12) 2097-2100
  • 18 Lippi G, Musa R, Avanzini P, Aloe R, Pipitone S, Sandei F. Influence of in vitro hemolysis on hematological testing on Advia 2120. Int J Lab Hematol 2012; 34 (2) 179-184
  • 19 Lippi G, Ippolito L, Fontana R. Prevalence of hemolytic specimens referred for arterial blood gas analysis. Clin Chem Lab Med 2011; 49 (5) 931-932
  • 20 Favaloro EJ. More on preanalytical variables affecting platelet function testing using light transmittance aggregometry. Clin Chem Lab Med 2011; 49 (4) 737-739
  • 21 Lippi G, Fontana R, Avanzini P , et al. Influence of mechanical trauma of blood and hemolysis on PFA-100 testing. Blood Coagul Fibrinolysis 2012; 23 (1) 82-86
  • 22 Bonar R, Favaloro EJ, Adcock DM. Quality in coagulation and haemostasis testing. Biochem Med (Zagreb) 2010; 20 (2) 184-199
  • 23 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 (6) 565-575
  • 24 Lippi G, Plebani M. Primary blood tubes mixing: time for updated recommendations. Clin Chem Lab Med 2012; 50 (4) 599-600
  • 25 Adcock Funk DM, Lippi G, Favaloro EJ. Quality standards for sample processing, transportation, and storage in hemostasis testing. Semin Thromb Hemost 2012; 38 (6) 576-585
  • 26 Lippi G, Plebani M. Continuous-flow automation and hemolysis index: a crucial combination. J Lab Autom 2012; (e-pub ahead of print). DOI: 10.1177/2211068212450014.
  • 27 Lippi G, Plebani M, Di Somma S, Cervellin G. Hemolyzed specimens: a major challenge for emergency departments and clinical laboratories. Crit Rev Clin Lab Sci 2011; 48 (3) 143-153
  • 28 Carraro P, Servidio G, Plebani M. Hemolyzed specimens: a reason for rejection or a clinical challenge?. Clin Chem 2000; 46 (2) 306-307
  • 29 Lippi G, Salvagno GL, Favaloro EJ, Guidi GC. Survey on the prevalence of hemolytic specimens in an academic hospital according to collection facility: opportunities for quality improvement. Clin Chem Lab Med 2009; 47 (5) 616-618
  • 30 Plebani M, Lippi G. Hemolysis index: quality indicator or criterion for sample rejection?. Clin Chem Lab Med 2009; 47 (8) 899-902
  • 31 Martin-Toutain I, Jobic L, Mancic T, Ankri A. [Evaluation of the automated coagulation analyzer Sysmex® CS-2100i (Siemens)]. Ann Biol Clin (Paris) 2011; 69 (6) 699-704
  • 32 Molenaar PJ, Leyte A. Pre-acquisition system assessment of the Sysmex® Coagulation System CS-2100i and comparison with end-user verification; a model for the regional introduction of new analysers and methods. Clin Chem Lab Med 2011; 49 (9) 1479-1489
  • 33 Lippi G. Interference studies: focus on blood cell lysates preparation and testing. Clin Lab 2012; 58 (3-4) 351-355
  • 34 Lippi G, Musa R, Aloe R, Mercadanti M, Pipitone S. Influence of temperature and period of freezing on the generation of hemolysate and blood cell lysate. Clin Biochem 2011; 44 (14–15) 1267-1269
  • 35 Dimeski G. Effects of hemolysis on the Roche ammonia method for Hitachi analyzers. Clin Chem 2004; 50 (5) 976-977
  • 36 Lippi G, Mercadanti M, Aloe R, Targher G. Erythrocyte mechanical fragility is increased in patients with type 2 diabetes. Eur J Intern Med 2012; 23 (2) 150-153
  • 37 Lippi G, Banfi G, Buttarello M , et al. Recommendations for detection and management of unsuitable samples in clinical laboratories. Clin Chem Lab Med 2007; 45 (6) 728-736
  • 38 Fraser CG, Lippi G, Plebani M. Reference change values may need some improvement but are invaluable tools in laboratory medicine. Clin Chem Lab Med 2012; 50 (5) 963-964
  • 39 Favaloro EJ, Adcock DM. Standardization of the INR: how good is your laboratory's INR and can it be improved?. Semin Thromb Hemost 2008; 34 (7) 593-603
  • 40 Lippi G, Favaloro EJ. Activated partial thromboplastin time: new tricks for an old dogma. Semin Thromb Hemost 2008; 34 (7) 604-611
  • 41 Lippi G, Cervellin G, Franchini M, Favaloro EJ. Biochemical markers for the diagnosis of venous thromboembolism: the past, present and future. J Thromb Thrombolysis 2010; 30 (4) 459-471
  • 42 Favaloro EJ, Lippi G, Franchini M. Contemporary platelet function testing. Clin Chem Lab Med 2010; 48 (5) 579-598
  • 43 Linden MD, Tran H, Woods R, Tonkin A. High platelet reactivity and antiplatelet therapy resistance. Semin Thromb Hemost 2012; 38 (2) 200-212
  • 44 Tripodi A. Problems and solutions for testing hemostasis assays while patients are on anticoagulants. Semin Thromb Hemost 2012; 38 (6) 586-592
  • 45 Walenga JM, Jackson CM, Kessler CM. Low molecular weight heparins differ substantially: impact on developing biosimilar drugs. Semin Thromb Hemost 2011; 37 (3) 322-327
  • 46 Lippi G, Franchini M, Favaloro EJ. Pharmacogenetics of vitamin K antagonists: useful or hype?. Clin Chem Lab Med 2009; 47 (5) 503-515
  • 47 Favaloro EJ, Lippi G. Laboratory testing and/or monitoring of the new oral anticoagulants/antithrombotics: for and against?. Clin Chem Lab Med 2011; 49 (5) 755-757
  • 48 Favaloro EJ, Lippi G, Koutts J. Laboratory testing of anticoagulants: the present and the future. Pathology 2011; 43 (7) 682-692
  • 49 Favaloro EJ, Lippi G. The new oral anticoagulants and the future of haemostasis laboratory testing. Biochem Med (Zagreb) 2012; 22 (3) 329-341
  • 50 Roß RS, Paar D. Analytisch und klinisch relevante Interferenzen in der Gerinnungsanalytik am Beispiel des MDA 180. Laboratoriumsmedizin 1998; 22: 90-94
  • 51 Laga AC, Cheves TA, Sweeney JD. The effect of specimen hemolysis on coagulation test results. Am J Clin Pathol 2006; 126 (5) 748-755
  • 52 Lippi G, Montagnana M, Salvagno GL, Guidi GC. Interference of blood cell lysis on routine coagulation testing. Arch Pathol Lab Med 2006; 130 (2) 181-184
  • 53 Lippi G, Avanzini P, Pavesi F , et al. Studies on in vitro hemolysis and utility of corrective formulas for reporting results on hemolyzed specimens. Biochem Med (Zagreb) 2011; 21 (3) 297-305
  • 54 Tantanate C, Teyateeti M, Tientadakul P. Influence of plasma interferences on screening coagulogram and performance evaluation of the automated coagulation analyzer Sysmex® CS-2100i. Siriraj Medical Journal 2011; 63 (5) 151-156
  • 55 Lippi G, Avanzini P, Zobbi V, Ippolito L. Influence of mechanical hemolysis of blood on two D-dimer immunoassays. Blood Coagul Fibrinolysis 2012; 23 (5) 461-463
  • 56 Bauer NB, Eralp O, Moritz A. Effect of hemolysis on canine kaolin-activated thromboelastography values and ADVIA 2120 platelet activation indices. Vet Clin Pathol 2010; 39 (2) 180-189
  • 57 Franchini M, Targher G, Lippi G. Serum bilirubin levels and cardiovascular disease risk: a Janus Bifrons?. Adv Clin Chem 2010; 50: 47-63
  • 58 Grafmeyer D, Bondon M, Manchon M, Levillain P. The influence of bilirubin, haemolysis and turbidity on 20 analytical tests performed on automatic analysers. Results of an interlaboratory study. Eur J Clin Chem Clin Biochem 1995; 33 (1) 31-52
  • 59 Dimeski G. Interference testing. Clin Biochem Rev 2008; 29 (Suppl. 01) S43-S48
  • 60 Quehenberger P, Kapiotis S, Handler S, Ruzicka K, Speiser W. Evaluation of the automated coagulation analyzer SYSMEX CA 6000. Thromb Res 1999; 96 (1) 65-71
  • 61 Appert-Flory A, Fischer F, Jambou D, Toulon P. Evaluation and performance characteristics of the automated coagulation analyzer ACL TOP. Thromb Res 2007; 120 (5) 733-743
  • 62 Fischer F, Appert-Flory A, Jambou D, Toulon P. Evaluation of the automated coagulation analyzer Sysmex CA-7000. Thromb Res 2006; 117 (6) 721-729
  • 63 Dorn-Beineke A, Dempfle CE, Bertsch T, Wisser H. Evaluation of the automated coagulation analyzer Sysmex CA-7000. Thromb Res 2005; 116 (2) 171-179
  • 64 Kroll MH. Evaluating interference caused by lipemia. Clin Chem 2004; 50 (11) 1968-1969
  • 65 Dimeski G, Jones BW. Lipaemic samples: effective process for lipid reduction using high speed centrifugation compared with ultracentrifugation. Biochem Med (Zagreb) 2011; 21 (1) 86-92
  • 66 Lippi G, Aloe R. Hyponatremia and pseudohyponatremia: first, do no harm. Am J Med 2010; 123 (9) e17
  • 67 Arambarri M, Oriol A, Sancho JM, Roncalés FJ, Galán A, Galimany R. [Interference in blood coagulation tests on lipemic plasma. Correction using n-hexane clearing]. Sangre (Barc) 1998; 43 (1) 13-19
  • 68 Bai B, Christie DJ, Gorman RT, Wu JR. Comparison of optical and mechanical clot detection for routine coagulation testing in a large volume clinical laboratory. Blood Coagul Fibrinolysis 2008; 19 (6) 569-576
  • 69 Lippi G, Salvagno GL, Lima-Oliveira G , et al. Interference of a regular standardized meal on routine coagulation testing. Poster presented at: Second Annual Joint Meeting SIBioC –SIMeL; October 27–30, 2009; Naples, Italy
  • 70 McEnroe RJ, Burritt MF, Powers DM, Rheinheimer DW. Interference Testing in Clinical Chemistry; Approved Guideline—Second Edition. Wayne, PA: Clinical Laboratory Standards Institute; 2005. . CLSI document EP7–A2
  • 71 Vermeer HJ, Steen G, Naus AJ, Goevaerts B, Agricola PT, Schoenmakers CH. Correction of patient results for Beckman Coulter LX-20 assays affected by interference due to hemoglobin, bilirubin or lipids: a practical approach. Clin Chem Lab Med 2007; 45 (1) 114-119