Haemostasis pp 73-83 | Cite as

Methods of Coagulation

  • Gemma Crighton
Part of the Methods in Molecular Biology book series (MIMB, volume 992)


The investigation of haemostasis is aimed to replicate normal blood coagulation in the laboratory. There are a number of different methods employed including clot-based coagulation tests, chromogenic assays, immunological tests, and aggregation studies. Today automated machines are designed to be able to perform a number of different methodologies.

To guarantee the results produced by a laboratory are reliable and accurate, laboratories need to follow international standards and guidelines and use established methodologies. The laboratory needs a quality management plan, to perform internal quality control and participate in an external quality assurance program.

Key words

Clot-based coagulation tests Chromogenic assays Immunological tests Aggregation studies Quality management Standardization 


  1. 1.
    McGlinchey K (2010) Coagulation automation. ADVANCE for Administrators of the Laboratory 19:26Google Scholar
  2. 2.
    Flanders MM, Crist R, Safapour S, Rodgers GM (2002) Evaluation and performance characteristics of the STA-R coagulation analyzer. Clin Chem 48:1622–1624PubMedGoogle Scholar
  3. 3.
    Milos M, Herak D, Kuric L, Horvat I, Zadro R (2007) Evaluation and performance characteristics of the coagulation system: ACL TOP analyzer—HemosIL reagents. Int J Lab Hematol 31:26–35CrossRefGoogle Scholar
  4. 4.
    Kitchen S, McCraw A, on behalf of the WFH Laboratory Sciences Committee (2010) Diagnosis of haemophilia and other bleeding disorders: a laboratory manual, 2nd edn. World Federation of Hemophilia, Montreal, QCGoogle Scholar
  5. 5.
    Lind SE (1991) The bleeding time does not predict surgical bleeding. Blood 77:2547–2552PubMedGoogle Scholar
  6. 6.
    Mielke CH Jr, Kaneshiro MM, Maher IA, Weiner JM, Rapaport SI (1969) The standardized normal Ivy bleeding time and its prolongation by aspirin. Blood 34:204–215PubMedGoogle Scholar
  7. 7.
    World Health Organisation (1997) Recommended methodology for using WHO International Preparations for thromboplastin, WHO, 1121 Geneva 27, SwitzerlandGoogle Scholar
  8. 8.
    Bai B, Christie DJ, Gorman RT, Wu JR (2008) Comparison of optical and mechanical clot detection for routine coagulation testing in a large volume clinical laboratory. Blood Coagul Fibrinolysis 19:569–576PubMedCrossRefGoogle Scholar
  9. 9.
    D’Angelo A, Seveso MP, D’Angelo SV, Gilardoni F, Macagni A, Manotti C, Bonini P (1989) Comparison of two automated coagulometers and the manual tilt-tube method for the determination of prothrombin time. Am J Clin Pathol 92:321–328PubMedGoogle Scholar
  10. 10.
    Johnstone IB (1984) The activated partial thromboplastin time of diluted plasma: variability due to the method of fibrin detection. Can J Comp Med 48:198–201PubMedGoogle Scholar
  11. 11.
    Lippi G, Franchini M, Favaloro EJ (2009) One-stage clotting versus chromogenic assays for assessing recombinant factor VIII: two faces of a haemostasis coin. Blood Coagul Fibrinolysis 20:1–3PubMedCrossRefGoogle Scholar
  12. 12.
    Chandler WL, Ferrell C, Lee J, Tun T, Kha H (2003) Comparison of three methods for measuring factor VIII levels in plasma. Am J Clin Pathol 120:34–39PubMedCrossRefGoogle Scholar
  13. 13.
    Keeling DM, Sukhu K, Kemball-Cook G, Waseem N, Bagnall R, Lloyd JV (1999) Diagnostic importance of the two-stage factor VIII:C assay demonstrated by a case of mild haemophilia associated with His 1954–> Leu substitution in the factor VIII A3 domain. Br J Haematol 105:1123–1126PubMedCrossRefGoogle Scholar
  14. 14.
    Fareed J, Messmore HL, Walenga JM, Bermes EW Jr (1983) Diagnostic efficacy of newer synthetic-substrates methods for assessing coagulation variables: a critical overview. Clin Chem 29:225–236PubMedGoogle Scholar
  15. 15.
    Hutton RA (1987) Chromogenic substrates in haemostasis. Blood Rev 1:201–206PubMedCrossRefGoogle Scholar
  16. 16.
    Svendsen L, Blomback B, Blomback M, Olson PI (1972) Synthetic chromogenic substrates for determination of trypsin, thrombin and thrombin-like enzymes. Thromb Res 1:267–278Google Scholar
  17. 17.
    Francis JL (1979) A new chromogenic assay for the specific determination of prothrombin. J Clin Pathol 32:651–654PubMedCrossRefGoogle Scholar
  18. 18.
    Tripodi A, Mannucci PM (1984) Clinical evaluation of a fully automated chromogenic method for prothrombin time compared with a conventional coagulation method. Clin Chem 30:1392–1395PubMedGoogle Scholar
  19. 19.
    Girolami A, Vettore S (2009) Discrepancies between clotting and chromogenic assays in congenital coagulation disorders: the hemostatic coin has only one face, chromogenic substrates many. Blood Coagul Fibrinolysis 20:484–485PubMedCrossRefGoogle Scholar
  20. 20.
    Peyvandi F, Menegatti M, Santagostino E, Akhavan S, Uprichard J, Perry DJ, Perkins SJ, Mannucci PM (2002) Gene mutations and three-dimensional structural analysis in 13 families with severe factor X deficiency. Br J Haematol 117:685–692PubMedCrossRefGoogle Scholar
  21. 21.
    Walker JM (ed) (1996) The protein protocols handbook. Humana, Totowa, NJGoogle Scholar
  22. 22.
    Palareti G, Maccaferri M, Manotti C, Tripodi A, Chantarangkul V, Rodeghiero F, Ruggeri M, Mannucci PM (1991) Fibrinogen assays: a collaborative study of six different methods. C.I.S.M.E.L. Comitato Italiano per la Standardizzazione dei Metodi in Ematologia e Laboratorio. Clin Chem 37:714–719PubMedGoogle Scholar
  23. 23.
    Verbruggen (1975) Quantitative immunoelectrophoretic methods: a literature survey. Clin Chem 21:5–43PubMedGoogle Scholar
  24. 24.
    Ruggeri ZM, Zimmerman TS (1981) The complex multimeric composition of factor VIII/von Willebrand factor. Blood 57:1140–1143PubMedGoogle Scholar
  25. 25.
    Engvall E (2010) The ELISA, enzyme-linked immunosorbent assay. Clin Chem 56:319–320PubMedCrossRefGoogle Scholar
  26. 26.
    Voller A, Bartlett A, Bidwell DE (1978) Enzyme immunoassays with special reference to ELISA techniques. J Clin Pathol 31:507–520PubMedCrossRefGoogle Scholar
  27. 27.
    Amiral J, Adalbert B, Adam M (1984) Application of enzyme immunoassays to coagulation testing. Clin Chem 30:1512–1516PubMedGoogle Scholar
  28. 28.
    van der Graaf F, van den Borne H, van der Kolk M, de Wild PJ, Janssen GW, van Uum SH (2000) Exclusion of deep venous thrombosis with D-dimer testing—comparison of 13 D-dimer methods in 99 outpatients suspected of deep venous thrombosis using venography as reference standard. Thromb Haemost 83:191–198PubMedGoogle Scholar
  29. 29.
    Pengo V, Thiagarajan P, Shapiro SS, Heine MJ (1987) Immunological specificity and mechanism of action of IgG lupus anticoagulants. Blood 70:69–76PubMedGoogle Scholar
  30. 30.
    Gella FS, Gener J, Latex J (1991) Agglutination procedures in immunodiagnosis. Pure Appl Chem 63:1131–1134CrossRefGoogle Scholar
  31. 31.
    Ewenstein BM (2001) Use of ristocetin cofactor activity in the management of von Willebrand disease. Haemophilia 7(Suppl 1):10–15PubMedCrossRefGoogle Scholar
  32. 32.
    Lewis SM, Bain BJ, Bates I (eds) (2006) Dacie and Lewis practical haematology, 10th edn. Churchill Livingstone Elsevier, Philadelphia, PAGoogle Scholar
  33. 33.
    Weiss HJ, Hoyer LW, Rickles FR, Varma A, Rogers J (1973) Quantitative assay of a plasma factor deficient in von Willebrand’s disease that is necessary for platelet aggregation. Relationship to factor VIII procoagulant activity and antigen content. J Clin Invest 52:2708–2716PubMedCrossRefGoogle Scholar
  34. 34.
    Born GV, Cross MJ (1963) The aggregation of blood platelets. J Physiol 168:178–195PubMedGoogle Scholar
  35. 35.
    Yardumian DA, Mackie IJ, Machin SJ (1986) Laboratory investigation of platelet function: a review of methodology. J Clin Pathol 39:701–712PubMedCrossRefGoogle Scholar
  36. 36.
    British Committee for Standards in Haematology (1988) Guidelines on platelet function testing. The British Society for Haematology BCSH Haemostasis and Thrombosis Task Force, 1988/12/01 ed, p 1322–1330, J Clin PatholGoogle Scholar
  37. 37.
    National Pathology Accreditation Advisory Council (2009) Requirements for participation in external quality assessment, Commonwealth of AustraliaGoogle Scholar
  38. 38.
    World Health Organisation (2011) International reference materials.
  39. 39.
    Hubbard AR, Kitchen S, Beeharry M, Bevan SA, Bowyer A, on behalf of the SSC Working Group on Coagulation Standards of the ISTH. (2011) Long-term stability of the Scientific and Standardization Committee Secondary Coagulation Standard (SSC Lot no. 3). J Thromb Haemost 9:2146–2148Google Scholar
  40. 40.
    Clinical Laboratory Standards Institute (2011) Working together to improve the quality of medical care–the Global Application of CLSI Documents, Lab Medicine, p 440–441Google Scholar
  41. 41.
    Lewis SM (1988) The WHO international external quality assessment scheme for haematology. Bull World Health Organ 66:283–290PubMedGoogle Scholar
  42. 42.
    Lewis SM (1996) Quality control: good laboratory practice—an overview of quality assurance. In: XXVIth World Congress of the International Society of Haematology. ICSH, SingaporeGoogle Scholar
  43. 43.
    Garvey MB, Black JM (1972) The detection of fibrinogen-fibrin degradation products by means of a new antibody-coated latex particle. J Clin Pathol 25:680–682PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press 2013

Authors and Affiliations

  • Gemma Crighton
    • 1
  1. 1.Department of HaematologyRoyal Children’s HospitalMelbourneAustralia

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