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Validation and standardization of the ETP-based activated protein C resistance test for the clinical investigation of steroid contraceptives in women: an unmet clinical and regulatory need

  • Jonathan Douxfils ORCID logo EMAIL logo , Laure Morimont ORCID logo , Anne-Sophie Delvigne , Philippe Devel , Bernard Masereel , Hélène Haguet , Céline Bouvy and Jean-Michel Dogné
Published/Copyright: August 24, 2019
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 58 Issue 2

Abstract

Background

Regulatory bodies recommend the use of an assay based on the assessment of the endogenous thrombin potential (ETP) for the investigation of the activated protein C resistance (APCr) in the development of steroid contraceptives in women. However, the assays described in the literature are home-made and not standardized regarding the method, the reagents, the reference plasma and the quality controls. In the absence of any commercially available method, we aimed at validating the ETP-based APCr assay.

Methods

The validation was performed according to regulatory standards. The method targets a 90% inhibition of the ETP in healthy donors in the presence of APC compared to the same condition in the absence of APC. As a large-scale production of a pool of plasma from well-selected healthy donors is impossible, algorithms were applied to a commercial reference plasma to correlate with the selected pool.

Results

Repeatability and intermediate precision passed the acceptance criteria. The assay demonstrated a curvilinear dose response to protein S and APC concentrations (R2 > 0.99). Analysis of plasma samples from 47 healthy individuals (22 women not taking combined hormonal contraceptives [CHC], and 25 men not Factor V Leiden carriers) confirmed the validity of the test, with a mean inhibition percentage of 90%. Investigations in 15 women taking different contraceptives and in two subjects with Factor V Leiden confirmed the good sensitivity and performance of the assay.

Conclusions

This validation provides the pharmaceutical industry, the regulatory bodies and physicians with a reproducible, sensitive and validated gold-standard ETP-based APCr assay.

Keywords: activated protein C resistance; blood coagulation test; combined hormonal contraceptive; endogenous thrombin potential; Factor V Leiden; protein S deficiency
Corresponding author: Prof. Jonathan Douxfils, Qualiblood sa, Namur, Belgium; and University of Namur, Department of Pharmacy, Namur Thrombosis and Hemostasis Center, NAmur Research Institute for LIfe Sciences, Namur, Belgium, Phone: +32 81 72 42 91, Fax: +32 81 72 42 99
aJonathan Douxfils, Laure Morimont, Céline Bouvy and Jean-Michel Dogné contributed equally as first and last authors, respectively.

Acknowledgments

The authors would like to thank François Mullier and the team from the Haematology Laboratory of the CHU UcL Namur, Yvoir, Belgium for providing support in Factor V Leiden investigations.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: The study was financed by QUALIblood and the Public Service of Wallonia (convention n° 1820173) in Belgium.

  3. Employment or leadership: Among the authors, J. Douxfils is CEO and founder of QUALIblood and reports personal fees from Diagnostica Stago, Roche, Roche Diagnostics, Portola and Daiichi-Sankyo, outside the submitted work.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(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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Received: 2019-05-06
Accepted: 2019-07-30
Published Online: 2019-08-24
Published in Print: 2020-01-28

©2019 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2019-0471
Keywords for this article
activated protein C resistance; blood coagulation test; combined hormonal contraceptive; endogenous thrombin potential; Factor V Leiden; protein S deficiency

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Towards a personalized assessment of vitamin D status
  4. Reviews
  5. Circulating tumor DNA and their added value in molecular oncology
  6. Telomere length determinants in childhood
  7. Opinion Papers
  8. Serum or plasma? An old question looking for new answers
  9. Evaluating sample stability in the clinical laboratory with the help of linear and non-linear regression analysis
  10. General Clinical Chemistry and Laboratory Medicine
  11. Simultaneous measurement of 25(OH)-vitamin D and 24,25(OH)2-vitamin D to define cut-offs for CYP24A1 mutation and vitamin D deficiency in a population of 1200 young subjects
  12. How well do Croatian laboratories adhere to national recommendations for laboratory diagnostics of chronic kidney disease (CKD)?
  13. Underfilling of vacuum blood collection tubes leads to increased lactate dehydrogenase activity in serum and heparin plasma samples
  14. Calcium state estimation by total calcium: the evidence to end the never-ending story
  15. The use of faecal immunochemical testing in the decision-making process for the endoscopic investigation of iron deficiency anaemia
  16. Measurement uncertainty of β-lactam antibiotics results: estimation and clinical impact on therapeutic drug monitoring
  17. Practical approach to method verification in plasma and validation in cerebrospinal fluid under accreditation using a flexible scope in molecular virology: setting up the HIV, HBV and HCV Aptima™ Quant Dx assays
  18. Plasma neurofilament light chain is associated with mortality after spontaneous intracerebral hemorrhage
  19. Comparison of the diagnostic performance of two automated urine sediment analyzers with manual phase-contrast microscopy
  20. Development and validation of LC-MS/MS methods to measure tobramycin and lincomycin in plasma, microdialysis fluid and urine: application to a pilot pharmacokinetic research study
  21. Reference Values and Biological Variations
  22. Cord blood S100B: reference ranges and interest for early identification of newborns with brain injury
  23. Hematology and Coagulation
  24. Validation and standardization of the ETP-based activated protein C resistance test for the clinical investigation of steroid contraceptives in women: an unmet clinical and regulatory need
  25. Cancer Diagnostics
  26. Next-generation sequencing for tumor mutation quantification using liquid biopsies
  27. Cardiovascular Diseases
  28. Evolution of the slopes of ST2 and galectin-3 during marathon and ultratrail running compared to a control group
  29. Letters to the Editor
  30. Biological variation of two serum markers for preeclampsia prediction
  31. Daily monitoring of a control material with a concentration near the limit of detection improves the measurement accuracy of highly sensitive troponin assays
  32. Are patients adequately informed about procedures for 24-h urine collection?
  33. Interferences in free thyroxine concentration using the Roche analytical platform: improvement of the third generation?
  34. Procedures for the diagnosis of macro-follicle stimulating hormone (FSH) in a patient with high serum FSH concentrations
  35. False-positive result of immunochromatographic (IC) strip test for the diagnosis of α-thalassemia in samples with autoantibodies
  36. Sample dilution for soluble interleukin-2 receptor α measurement: comparison of two different matrices
  37. The growing problem of predatory publishing: a case report
  38. Cerebrospinal fluid lactate levels according to the site of puncture
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