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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter

IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
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Volume 57, Issue 4


Next-generation rapid serum tube technology using prothrombin activator coagulant: fast, high-quality serum from normal samples

Kong-Nan Zhao
  • Centre for Venomics Research, School of Medicine, The University of Queensland, Translational Research Institute, Woolloongabba, QLD, Australia
  • Centre for Clinical Research, The University of Queensland, Royal Brisbane and Women’s Hospital Campus, Herston, Brisbane, QLD, Australia
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Goce Dimeski
  • Chemical Pathology, Princess Alexandra Hospital, Woolloongabba, Brisbane, QLD, Australia
  • School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD, Australia
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ John de Jersey
  • School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD, Australia
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Lambro A. Johnson
  • Centre for Venomics Research, School of Medicine, The University of Queensland, Translational Research Institute, Woolloongabba, QLD, Australia
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Michael Grant / Paul P. Masci
  • Centre for Venomics Research, School of Medicine, The University of Queensland, Translational Research Institute, Woolloongabba, QLD, Australia
  • Centre for Clinical Research, The University of Queensland, Royal Brisbane and Women’s Hospital Campus, Herston, Brisbane, QLD, Australia
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Martin F. Lavin
  • Corresponding author
  • Centre for Clinical Research, The University of Queensland, Royal Brisbane and Women’s Hospital Campus, Herston, Brisbane, QLD 4029, Australia, Phone: +61 07 3346 6045, Fax: +61 07 3346 5509
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2018-09-29 | DOI: https://doi.org/10.1515/cclm-2018-0397



Incomplete blood clotting or latent clotting in serum is a common laboratory problem, especially for patients on anticoagulant therapy or when serum tubes are centrifuged before clotting is completed. We describe a novel approach to producing high-quality serum using snake venom prothrombin activator complex (OsPA) as an additive in blood collection tubes for non-anticoagulated (normal) individuals.


Plasma clotting assays were performed using a Hyland-Clotek instrument. Blood clotting was visually observed, and thromboelastography was also performed to determine the important parameters of coagulation. Thrombin generation was assayed using the chromogenic substrate S-2238, and biochemical analytes in the serum were determined on chemistry and immunoassay analysers. Fibrinogen was determined by either ELISA or Clauss fibrinogen assay.


We initially showed that OsPA had strong coagulation activity in clotting not only recalcified citrated plasma and recalcified citrated whole blood, but also fresh whole blood in a clinical setting. The use of TEG clearly showed improved speed of clotting and generation of a firmer clot. We also showed that the use of OsPA to produce serum did not interfere with the determination of commonly measured biochemical analytes. The underlying clotting mechanism involves a burst of thrombin production at the initial stages of the clotting process upon contact with prothrombin in blood.


These results demonstrate rapid generation of high-quality serum, contributing to faster turnaround times with standardised quality samples, for accurate analyte determinations in normal individuals.

This article offers supplementary material which is provided at the end of the article.

Keywords: analyte determination; blood clotting; high-quality serum; prothrombin activators; snake venom proteins; thrombin generation


  • 1.

    Vignati G, Chiecchio A, Osnaghi B, Giovanelli L, Meloncelli C. Different biological matrices (serum and plasma) utilization in consolidation processes: evaluation of seven access immunoassays. Clin Chem Lab Med 2008;46:264–70.PubMedWeb of ScienceGoogle Scholar

  • 2.

    Lee BS, Lee YU, Kim HS, Kim TH, Park J, Lee JG, et al. Fully integrated lab-on-a-disc for simultaneous analysis of biochemistry and immunoassay from whole blood. Lab Chip 2011;11:70–8.Web of ScienceCrossrefPubMedGoogle Scholar

  • 3.

    Bowen RA, Sattayapiwat A, Gounden V, Remaley AT. Blood collection tube-related alterations in analyte concentrations in quality control material and serum specimens. Clin Biochem 2014;47:150–7.CrossrefWeb of SciencePubMedGoogle Scholar

  • 4.

    Aoki N, Sakata Y, Ichinose A. Fibrin-associated plasminogen activation in alpha 2-plasmin inhibitor deficiency. Blood 1983;62:1118–22.PubMedGoogle Scholar

  • 5.

    Cuhadar S, Atay A, Koseoglu M, Dirican A, Hur A. Stability studies of common biochemical analytes in serum separator tubes with or without gel barrier subjected to various storage conditions. Biochem Med (Zagreb) 2012;22:202–14.PubMedGoogle Scholar

  • 6.

    Dimeski G, Masci PP, Trabi M, Lavin MF, De Jersey J. Evaluation of the Becton-Dickinson rapid serum tube: does it provide a suitable alternative to lithium heparin plasma tubes? Clin Chem Lab Med 2010;48:651–7.PubMedWeb of ScienceGoogle Scholar

  • 7.

    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.Web of ScienceCrossrefPubMedGoogle Scholar

  • 8.

    Bowen RA, Remaley AT. Interferences from blood collection tube components on clinical chemistry assays. Biochem Med (Zagreb) 2014;24:31–44.PubMedGoogle Scholar

  • 9.

    Prusa R, Doupovcova J, Warunek D, Stankovic AK. Improving laboratory efficiencies through significant time reduction in the preanalytical phase. Clin Chem Lab Med 2010;48:293–6.PubMedWeb of ScienceGoogle Scholar

  • 10.

    Dimeski G, Johnston J, Masci PP, Zhao KN, Brown N. Evaluation of the Greiner Bio-One serum separator BCA fast clot tube. Clin Chem Lab Med 2017;55:1135–41.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 11.

    Bompiani KM, Lohrmann JL, Pitoc GA, Frederiksen JW, Mackensen GB, Sullenger BA. Probing the coagulation pathway with aptamers identifies combinations that synergistically inhibit blood clot formation. Chem Biol 2014;21:935–44.PubMedWeb of ScienceCrossrefGoogle Scholar

  • 12.

    Brummel-Ziedins K, Undas A, Orfeo T, Gissel M, Butenas S, Zmudka K, et al. Thrombin generation in acute coronary syndrome and stable coronary artery disease: dependence on plasma factor composition. J Thromb Haemost 2008;6:104–10.PubMedGoogle Scholar

  • 13.

    Ruggeri ZM, Ruggeri ZM. Platelet and von Willebrand factor interactions at the vessel wall. Hamostaseologie 2004;24:1–11.CrossrefPubMedGoogle Scholar

  • 14.

    Undas A, Szuldrzynski K, Brummel-Ziedins KE, Tracz W, Zmudka K, Mann KG. Systemic blood coagulation activation in acute coronary syndromes. Blood 2009;113:2070–8.CrossrefWeb of SciencePubMedGoogle Scholar

  • 15.

    Stubbs MT, Bode W. Coagulation factors and their inhibitors. Curr Opin Struct Biol 1994;4:823–32.CrossrefPubMedGoogle Scholar

  • 16.

    Matafonov A, Sarilla S, Sun MF, Sheehan JP, Serebrov V, Verhamme IM, et al. Activation of factor Xi by products of prothrombin activation. Blood 2011;118:437–45.CrossrefWeb of SciencePubMedGoogle Scholar

  • 17.

    Walker FJ, Owen WG, Esmon CT. Characterization of the prothrombin activator from the venom of Oxyuranus scutellatus scutellatus (Taipan Venom). Biochemistry 1980;19:1020–3.PubMedCrossrefGoogle Scholar

  • 18.

    Kini RM. The intriguing world of prothrombin activators from snake venom. Toxicon 2005;45:1133–45.PubMedCrossrefGoogle Scholar

  • 19.

    Speijer H, Govers-Riemslag JW, Zwaal RF, Rosing J. Prothrombin activation by an activator from the venom of Oxyuranus scutellatus (Taipan Snake). J Biol Chem 1986;261:13258–67.PubMedGoogle Scholar

  • 20.

    Speijer H, Govers-Riemslag JW, Zwaal RF, Rosing J. Platelet procoagulant properties studied with snake venom prothrombin activators. Thromb Haemost 1987;57:349–55.PubMedCrossrefGoogle Scholar

  • 21.

    Bos MH, Camire RM. Procoagulant adaptation of a blood coagulation prothrombinase-like enzyme complex in Australian elapid venom. Toxins (Basel) 2010;2:1554–67.CrossrefPubMedGoogle Scholar

  • 22.

    Bos MH, Camire RM. Blood coagulation factors V and VIII: molecular mechanisms of procofactor activation. J Coagul Disord 2010;2:19–27.PubMedGoogle Scholar

  • 23.

    Austen DE, Rhymes IL. Laboratory Manual of Blood Coagulation. Oxford: Blackwell Scientific Publications, 1975:109.Google Scholar

  • 24.

    Swallow RA, Agarwala RA, Dawkins KD, Curzen NP. Thromboelastography: potential bedside tool to assess the effects of antiplatelet therapy? Platelets 2006;17:385–92.CrossrefPubMedGoogle Scholar

  • 25.

    Masci PP, Whitaker AN, De Jersey J. Purification and characterization of a prothrombin activator from the venom of the Australian brown snake, Pseudonaja textilis textilis. Biochem Int 1988;17:825–35.PubMedGoogle Scholar

  • 26.

    Moore GW, Tugnait S, Savidge GF. A new-generation dilute Russell’s viper venom time assay system for lupus anticoagulants: evaluation of detection utilising frozen reagents and controls. Br J Biomed Sci 2005;62:127–32.PubMedCrossrefGoogle Scholar

  • 27.

    Nowak G. The Ecarin clotting time, a universal method to quantify direct thrombin inhibitors. Pathophysiol Haemost Thromb 2003;33:173–83.PubMedCrossrefGoogle Scholar

  • 28.

    Van Cott EM, Smith EY, Galanakis DK. Elevated fibrinogen in an acute phase reaction prolongs the reptilase time but typically not the thrombin time. Am J Clin Pathol 2002;118: 263–8.CrossrefGoogle Scholar

  • 29.

    Herrera M, Paiva OK, Pagotto AH, Segura A, Serrano SM, Vargas M, et al. Antivenomic characterization of two antivenoms against the venom of the Taipan, Oxyuranus scutellatus, from Papua New Guinea and Australia. Am J Trop Med Hyg 2014;91:887–94.Web of ScienceCrossrefPubMedGoogle Scholar

  • 30.

    Arslan FD, Karakoyun I, Basok BI, Aksit MZ, Baysoy A, Ozturk YK, et al. The local clinical validation of a new lithium heparin tube with a barrier: Bd Vacutainer(R) Barricor Lh Plasma tube. Biochem Med (Zagreb) 2017;27:030706.PubMedCrossrefGoogle Scholar

  • 31.

    Carraro P, Servidio G, Plebani M. Hemolyzed specimens: a reason for rejection or a clinical challenge? Clin Chem 2000;46:306–7.Google Scholar

  • 32.

    Lippi G, Cervellin G, Mattiuzzi C. Critical review and meta-analysis of spurious hemolysis in blood samples collected from intravenous catheters. Biochem Med (Zagreb) 2013;23:193–200.PubMedGoogle Scholar

  • 33.

    Monneret D, Mestari F, Atlan G, Corlouer C, Ramani Z, Jaffre J, et al. Hemolysis indexes for biochemical tests and immunoassays on roche analyzers: determination of allowable interference limits according to different calculation methods. Scand J Clin Lab Invest 2015;75:162–9.Web of SciencePubMedCrossrefGoogle Scholar

  • 34.

    Rao VS, Kini RM. Pseutarin C, a prothrombin activator from Pseudonaja textilis venom: its structural and functional similarity to mammalian coagulation factor Xa-Va complex. Thromb Haemost 2002;88:611–9.CrossrefPubMedGoogle Scholar

  • 35.

    St Pierre L, Masci PP, Filippovich I, Sorokina N, Marsh N, Miller DJ, et al. Comparative analysis of prothrombin activators from the venom of Australian Elapids. Mol Biol Evol 2005;22:1853–64.CrossrefPubMedGoogle Scholar

  • 36.

    Bos MH, Boltz M, St Pierre L, Masci PP, De Jersey J, Lavin MF, et al. Venom Factor V from the common brown snake escapes hemostatic regulation through procoagulant adaptations. Blood 2009;114:686–92.Web of SciencePubMedCrossrefGoogle Scholar

  • 37.

    Plebani M, Lippi G. Improving diagnosis and reducing diagnostic errors: the next frontier of laboratory medicine. Clin Chem Lab Med 2016;54:1117–8.Web of SciencePubMedGoogle Scholar

  • 38.

    Lippi G, Plebani M, Favaloro EJ. The model list of essential in vitro diagnostics: nuisance or opportunity? Diagnosis (Berl) 2018. pii: /j/dx.ahead-of-print/dx-2018-0035/dx-2018-0035.xml. Doi: 10.1515/dx-2018-0035. [Epub ahead of print].PubMed

  • 39.

    Plebani M, Lippi G. Improving the post-analytical phase. Clin Chem Lab Med 2010;48:435–6.Web of SciencePubMedGoogle Scholar

About the article

Received: 2018-04-17

Accepted: 2018-08-31

Published Online: 2018-09-29

Published in Print: 2019-03-26

Author contributions: M.F.L, J.D.J., P.P.M., G.D., K.N.Z. and M.G. contributed to the conception of this project and designed the experiments. P.P.M., K.N.Z., L.A.J. and G.D. performed the experiments. K.N.Z. and G.D. organised and analysed data, and all authors reviewed and discussed. K.N.Z. and M.F.L wrote the paper with input from all authors on the drafting of the manuscript. 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: MFL, PM, JdJ, MG and GD have Employee Share Options Scheme in Q-Sera. 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.

Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 57, Issue 4, Pages 483–497, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2018-0397.

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