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.
We wish to thank the Australian Red Cross Blood Supply (ARCBS) for the provision of blood for some experiments. The project was in part funded by a development grant from the National Health and Medical Research Council of Australia and by Q-Sera Pty Ltd., Australia, Grant Number: Clotting tube product development. We wish to thank Nathan Dunstan, Venom Supplies, South Australia, for providing snake venom and Ms Connie Solano, Haematology Department, Princess Alexandra Hospital, Brisbane, for supply of blood.
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.
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The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2018-0397).
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