Home Differences in lupus anticoagulant final conclusion through clotting time or Rosner index for mixing test interpretation
Article
Licensed
Unlicensed Requires Authentication

Differences in lupus anticoagulant final conclusion through clotting time or Rosner index for mixing test interpretation

  • Barbara Depreter and Katrien M.J. Devreese EMAIL logo
Published/Copyright: February 19, 2016
Become an author with De Gruyter Brill
Submit Manuscript Author Information
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 54 Issue 9

Abstract

Background:

Lupus anticoagulant (LAC) testing includes a screening, mixing and confirmation step. Although recently published guidelines on LAC testing are a useful step towards standardization, a lack of consensus remains whether to express mixing tests in clotting time (CT) or index of circulating anticoagulant (ICA). The influence of anticoagulant therapy, e.g. vitamin K antagonists (VKA) or direct oral anticoagulants (DOAC) on both methods of interpretation remains to be investigated. The objective of this study was to contribute to a simplification and standardization of the LAC three-step interpretation on the level of the mixing test.

Methods:

Samples from 148 consecutive patients with LAC request and prolonged screening step, and 77 samples from patients non-suspicious for LAC treated with VKA (n=37) or DOAC (n=30) were retrospectively evaluated. An activated partial thromboplastin time (aPTT) and dilute Russell’s viper venom time (dRVVT) were used for routine LAC testing. The supplemental anticoagulant samples were tested with dRVVT only. We focused on the interpretation differences for mixing tests expressed as CT or ICA and compared the final LAC conclusion within each distinct group of concordant and discordant mixing test results.

Results:

Mixing test interpretation by CT resulted in 10 (dRVVT) and 16 (aPTT) more LAC positive patients compared to interpretation with ICA. Isolated prolonged dRVVT screen mix ICA results were exclusively observed in samples from VKA-treated patients without suspicion for LAC.

Conclusions:

We recommend using CT in respect to the 99th percentile cut-off for interpretation of mixing steps in order to reach the highest sensitivity and specificity in LAC detection.

Keywords: anticoagulant drugs; clotting time; lupus anticoagulant; mixing test; Rosner index
Corresponding author: Prof. Dr. Katrien M.J. Devreese, Coagulation Laboratory, Department of Clinical Chemistry, Microbiology and Immunology, Ghent University Hospital, De Pintelaan 185 (2P8), 9000 Ghent, Belgium, Phone: +32 9 332 65 67, Fax: +32 9 332 49 85, E-mail:

Acknowledgments

The authors thank Michael Luypaert, An De Saar and Fien Matthys for LAC analyses of the samples.

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

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  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.

References

1. Brandt JT, Triplett DA, Alving B, Scharrer I. Criteria for the diagnosis of lupus anticoagulants – an update. Thromb Haemost 1995;74:1185–90.10.1055/s-0038-1649901Search in Google Scholar

2. Machin SJ, Giddings JC, Greaves M, Hutton RA, Mackie IJ, Malia RG, et al. Guidelines on testing for the lupus anticoagulant. J Clin Pathol 1991;44:885–9.Search in Google Scholar

3. Pengo V, Tripodi A, Reber G, Rand JH, Ortel TL, Galli M, et al. Update of the guidelines for lupus anticoagulant detection. Subcommittee on Lupus Anticoagulant/Antiphospholipid Antibody of the Scientific and Standardisation Committee of the International Society on Thrombosis and Haemostasis. J Thromb Haemost 2009;7:1737–40.10.1111/j.1538-7836.2009.03555.xSearch in Google Scholar PubMed

4. Tripodi A, Pengo V. More on: laboratory investigation of lupus anticoagulants: mixing studies are sometimes required. J Thromb Haemost 2011;9:2126–7.10.1111/j.1538-7836.2011.04440.xSearch in Google Scholar PubMed

5. Devreese KM. No more mixing tests required for integrated assay systems in the laboratory diagnosis of lupus anticoagulants? J Thromb Haemost 2010;8:1120–2.10.1111/j.1538-7836.2010.03807.xSearch in Google Scholar PubMed

6. Favaloro EJ, Bonar R, Zebeljan D, Kershaw G, Marsden K. Laboratory investigation of lupus anticoagulants: mixing studies are sometimes required. Thromb Haemost 2010;8:2828–31.10.1111/j.1538-7836.2010.04090.xSearch in Google Scholar PubMed

7. Chandler JB, Torres R, Rinder HM, Tormey CA. Lupus anticoagulant testing and anticoagulation do not mix: quantitation of discrepant results and potential approaches to reduce false positives. Br J Haematol 2014;167:704–7.10.1111/bjh.13030Search in Google Scholar PubMed PubMed Central

8. Pennings MT, De Groot PG, Meijers JC, Huisman A, Derksen RH, Urbanus RT. Optimisation of lupus anticoagulant tests: should test samples always be mixed with normal plasma? Thromb Haemost 2014;112:736–42.10.1160/TH14-02-0122Search in Google Scholar PubMed

9. Aboud M, Morel-Kopp MC, Ward C, Coyle L. False-negative or false-positive: laboratory diagnosis of lupus anticoagulant at the time of commencement of anticoagulant. J Thromb Haemost 2010;8:2070–3.10.1111/j.1538-7836.2010.03946.xSearch in Google Scholar PubMed

10. Aboud M, Morel-Kopp MC, Ward C, Coyle L. False negative or false positive: laboratory diagnosis of lupus anticoagulant at the time of commencement of anticoagulant: reply to a rebuttal. J Thromb Haemost 2011;9:1436–7.10.1111/j.1538-7836.2011.04338.xSearch in Google Scholar

11. Devreese KM, de Laat B. Mixing studies in lupus anticoagulant testing are required at least in some type of samples. J Thromb Haemost 2015;13:1475–8.10.1111/jth.13020Search in Google Scholar PubMed

12. Clinical and Laboratory Standards Institute. Laboratory testing for the lupus anticoagulant; Approved guideline, Wayne: CLSI Document H60-A, 2014. 2014.Search in Google Scholar

13. Moore GW, Savidge GF. The dilution effect of equal volume mixing studies compromises confirmation of inhibition by lupus anticoagulants even when mixture specific reference ranges are applied. Thromb Res 2006;118:523–8.10.1016/j.thromres.2005.09.010Search in Google Scholar PubMed

14. Chang SH, Tillema V, Scherr D. A “percent correction” formula for evaluation of mixing studies. Am J Clin Pathol 2002;117:62–73.10.1309/RREK-8L6M-D2KC-HWLHSearch in Google Scholar PubMed

15. Devreese KM. Interpretation of normal plasma mixing studies in the laboratory diagnosis of lupus anticoagulants. Thromb Res 2007;119:369–76.10.1016/j.thromres.2006.03.012Search in Google Scholar PubMed

16. Rosner E, Pauzner R, Lusky A, Modan M, Many A. Detection and quantitative evaluation of lupus circulating anticoagulant activity. Thromb Haemost 1987;57:144–7.10.1055/s-0038-1651083Search in Google Scholar

17. Triplett DA, Barna LK, Unger GA. A hexagonal (II) phase phospholipid neutralization assay for lupus anticoagulant identification. Thromb Haemost 1993;70:787–93.10.1055/s-0038-1649671Search in Google Scholar

18. Triplett DA. Assays for detection of antiphospholipid antibodies. Lupus 1994;3:281–7.10.1177/096120339400300415Search in Google Scholar PubMed

19. Tripodi A. Laboratory testing for lupus anticoagulants: a review of issues affecting results. Clin Chem 2007;53:1629–35.10.1373/clinchem.2007.089524Search in Google Scholar PubMed

20. Martinuzzo ME, Cerrato GS, Varela ML, Adamczuk YP, Forastiero RR. New guidelines for lupus anticoagulant: sensitivity and specificity of cut-off values calculated with plasmas from healthy controls in mixing and confirmatory tests. Int J Lab Hem 2012;34:208–13.10.1111/j.1751-553X.2011.01376.xSearch in Google Scholar PubMed

21. Kumano O, Ieko M, Naito S, Yoshida M, Takahashi N, Suzuki T, et al. Verification of the guidelines for lupus anticoagulant detection: Usefulness of index for circulating anticoagulant in APTT mixing test. Thromb Res 2014;134:503–9.10.1016/j.thromres.2014.05.030Search in Google Scholar PubMed

22. Dembitzer FR, Ledford Kraemer MR, Meijer P, Peerschke EI. Lupus anticoagulant testing: performance and practices by north american clinical laboratories. Am J Clin Pathol 2010;134:764–73.10.1309/AJCP4SPPLG5XVIXFSearch in Google Scholar PubMed

23. Arachchillage DR, Mackie IJ, Efthymiou M, Isenberg DA, Machin SJ, Cohen H. Interactions between rivaroxaban and antiphospholipid antibodies in thrombotic antiphospholipid syndrome. J Thromb Haemost 2015;13:1264–73.10.1111/jth.12917Search in Google Scholar PubMed

24. Martinuzzo ME, Barrera LH, D’adamo MA, Otaso JC, Gimenez MI, Oyhamburu J. Frequent false-positive results of lupus anticoagulant tests in plasmas of patients receiving the new oral anticoagulants and enoxaparin. Int J Lab Hem 2014;36:144–50.10.1111/ijlh.12138Search in Google Scholar PubMed

25. van Os GM, de Laat B, Kamphuisen PW, Meijers JC, de Groot PG. Detection of lupus anticoagulant in the presence of rivaroxaban using Taipan snake venom time. J Thromb Haemost 2011;9:1657–9.10.1111/j.1538-7836.2011.04395.xSearch in Google Scholar PubMed

Supplemental Material:

The online version of this article (DOI: 10.1515/cclm-2015-0978) offers supplementary material, available to authorized users.

Received: 2015-10-8
Accepted: 2016-1-18
Published Online: 2016-2-19
Published in Print: 2016-9-1

©2016 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Editorials
  3. The Theranos phenomenon, scientific transparency and freedom of speech
  4. Holotranscobalamin: in the middle of difficultly lies opportunity
  5. Review
  6. Laboratory and clinical risk assessment to treat myelodysplatic syndromes
  7. Mini Review
  8. Quantitative nucleic acid amplification by digital PCR for clinical viral diagnostics
  9. Genetics and Molecular Diagnostics
  10. Hybrid minigene splicing assay verified the pathogenicity of a novel splice site variant in the dystrophin gene of a Chinese patient with typical Duchenne muscular dystrophy phenotype
  11. General Clinical Chemistry and Laboratory Medicine
  12. Prospective validation of an automated chemiluminescence-based assay of renin and aldosterone for the work-up of arterial hypertension
  13. Sex steroid hormone stability in serum tubes with and without separator gels
  14. Reduced absorption and enhanced synthesis of cholesterol in patients with cystic fibrosis: a preliminary study of plasma sterols
  15. An International Standard for holotranscobalamin (holoTC): international collaborative study to assign a holoTC value to the International Standard for vitamin B12 and serum folate
  16. A technical and clinical evaluation of a new assay for inhibin A and its use in second trimester Down syndrome screening
  17. Investigation on the ability of first trimester glycodelin and angiopoietin-2 to predict small-for-gestational age pregnancies at delivery
  18. Plasma total C-terminal agrin fragment (tCAF) as a marker for kidney function in patients with chronic kidney disease
  19. Hematology and Coagulation
  20. Thirty-minutes’ exposure to smartphone call triggers neutrophil activation in vitro
  21. Performance of the XN-2000 WPC channel-flagging to differentiate reactive and neoplastic leukocytosis
  22. Differences in lupus anticoagulant final conclusion through clotting time or Rosner index for mixing test interpretation
  23. Reference Values and Biological Variations
  24. Derivation of level-specific reference change values (RCV) from a health screening database and optimization of their thresholds based on clinical utility
  25. Cancer Diagnosis
  26. BRAF analysis before surgery for papillary thyroid carcinoma: correlation with clinicopathological features and prognosis in a single-institution prospective experience
  27. Letters to the Editor
  28. Significant increase of serum prostate-specific antigen after exercise
  29. Serum delipidation but not high-speed centrifugation is effective in clearing lipemia interference in serum lipase activity measurement
  30. A relationship between absolute monocyte count and C-reactive protein in patients with migraine undergoing no pharmacological therapy
  31. Validation of the “Vacutainer® urinalysis preservative plus urine tube” for the determination of albumin and protein
  32. Performance evaluation of a novel automated HIV Ag/Ab chemiluminescence immunoassay
  33. SLC26A4 genotypes associated with enlarged vestibular aqueduct malformation in south Italian children with sensorineural hearing loss
  34. Is cystatin C level altered in women with polycystic ovary syndrome?
  35. Venous thromboembolism in a patient with persistent inhibitor to coagulation factor V – a case report
  36. Free light chains nephelometric assay: human urine stability in different storage conditions
  37. Comparing the viscoelastomeric fibrin polymerization assays FIBTEM® (ROTEM) vs. Functional Fibrinogen® (TEG): or why is a higher threshold for fibrinogen substitution better than a lower one?
  38. Reply to: Comparing the visco-elastomeric fibrin polymerization assays FIBTEM® (ROTEM) vs. Functional Fibrinogen® (TEG): or why is a higher threshold for fibrinogen substitution better than a lower one? By Schöchl et al.
  39. Congress Abstracts
  40. 5th Slovenian Congress of Clinical Chemistry and Laboratory Medicine
https://doi.org/10.1515/cclm-2015-0978
Keywords for this article
anticoagulant drugs; clotting time; lupus anticoagulant; mixing test; Rosner index

Articles in the same Issue

  1. Frontmatter
  2. Editorials
  3. The Theranos phenomenon, scientific transparency and freedom of speech
  4. Holotranscobalamin: in the middle of difficultly lies opportunity
  5. Review
  6. Laboratory and clinical risk assessment to treat myelodysplatic syndromes
  7. Mini Review
  8. Quantitative nucleic acid amplification by digital PCR for clinical viral diagnostics
  9. Genetics and Molecular Diagnostics
  10. Hybrid minigene splicing assay verified the pathogenicity of a novel splice site variant in the dystrophin gene of a Chinese patient with typical Duchenne muscular dystrophy phenotype
  11. General Clinical Chemistry and Laboratory Medicine
  12. Prospective validation of an automated chemiluminescence-based assay of renin and aldosterone for the work-up of arterial hypertension
  13. Sex steroid hormone stability in serum tubes with and without separator gels
  14. Reduced absorption and enhanced synthesis of cholesterol in patients with cystic fibrosis: a preliminary study of plasma sterols
  15. An International Standard for holotranscobalamin (holoTC): international collaborative study to assign a holoTC value to the International Standard for vitamin B12 and serum folate
  16. A technical and clinical evaluation of a new assay for inhibin A and its use in second trimester Down syndrome screening
  17. Investigation on the ability of first trimester glycodelin and angiopoietin-2 to predict small-for-gestational age pregnancies at delivery
  18. Plasma total C-terminal agrin fragment (tCAF) as a marker for kidney function in patients with chronic kidney disease
  19. Hematology and Coagulation
  20. Thirty-minutes’ exposure to smartphone call triggers neutrophil activation in vitro
  21. Performance of the XN-2000 WPC channel-flagging to differentiate reactive and neoplastic leukocytosis
  22. Differences in lupus anticoagulant final conclusion through clotting time or Rosner index for mixing test interpretation
  23. Reference Values and Biological Variations
  24. Derivation of level-specific reference change values (RCV) from a health screening database and optimization of their thresholds based on clinical utility
  25. Cancer Diagnosis
  26. BRAF analysis before surgery for papillary thyroid carcinoma: correlation with clinicopathological features and prognosis in a single-institution prospective experience
  27. Letters to the Editor
  28. Significant increase of serum prostate-specific antigen after exercise
  29. Serum delipidation but not high-speed centrifugation is effective in clearing lipemia interference in serum lipase activity measurement
  30. A relationship between absolute monocyte count and C-reactive protein in patients with migraine undergoing no pharmacological therapy
  31. Validation of the “Vacutainer® urinalysis preservative plus urine tube” for the determination of albumin and protein
  32. Performance evaluation of a novel automated HIV Ag/Ab chemiluminescence immunoassay
  33. SLC26A4 genotypes associated with enlarged vestibular aqueduct malformation in south Italian children with sensorineural hearing loss
  34. Is cystatin C level altered in women with polycystic ovary syndrome?
  35. Venous thromboembolism in a patient with persistent inhibitor to coagulation factor V – a case report
  36. Free light chains nephelometric assay: human urine stability in different storage conditions
  37. Comparing the viscoelastomeric fibrin polymerization assays FIBTEM® (ROTEM) vs. Functional Fibrinogen® (TEG): or why is a higher threshold for fibrinogen substitution better than a lower one?
  38. Reply to: Comparing the visco-elastomeric fibrin polymerization assays FIBTEM® (ROTEM) vs. Functional Fibrinogen® (TEG): or why is a higher threshold for fibrinogen substitution better than a lower one? By Schöchl et al.
  39. Congress Abstracts
  40. 5th Slovenian Congress of Clinical Chemistry and Laboratory Medicine
Sign up now to receive a 20% welcome discount
Institutional Access
How does access work?
Have an idea on how to improve our website?
Please write us.
© 2025 De Gruyter Brill
Downloaded on 17.9.2025 from https://www.degruyterbrill.com/document/doi/10.1515/cclm-2015-0978/html
Scroll to top button