Newly developed dilute Russell’s viper venom reagents for lupus anticoagulant detection with improved specificity

Abstract

Background

Dilute Russell’s viper venom time (dRVVT) is indispensible in lupus anticoagulant (LA) detection yet commercial reagents from different suppliers perform variably, no gold standard assays exist and therapeutic anticoagulation interference is problematic.

Objective

The objective of this study was to compare a new formulation dRVVT with two currently available dRVVTs.

Materials and methods

Life Diagnostics (LD) dRVVT and Stago PTT-LA were routinely used for lupus anticoagulant detection, plus Taipan snake venom time/ecarin time (TSVT/ET) for patients on warfarin or rivaroxaban. Siemens dRVVT and the new HYPHEN BioMed (HBM) dRVVT were tested with 193 patient samples. Group 1, 59 non-anticoagulated patients (NAPs) LA-positive in LD dRVVT; Group 2, 15 PTT-LA-positive/dRVVT-negative NAPs; Group 3, 24 LA-positive warfarinized patients; Group 4, 13 patients on rivaroxaban; Group 5, 62 LA-negative thrombotic NAPs; Group 6, 20 warfarinized, non-antiphospholipid syndrome patients.

Results

Accepting that the Life Diagnostics reagents were acting as a pseudo-gold standard, Siemens dRVVT detected 56/59, (95%) Group 1 LA and HBM dRVVT 46/59, (76%), one each from Group 2, and Siemens dRVVT detected one in Group 5. The lower HBM dRVVT detection rate mainly concerned weaker LA, where between-reagent concordance is problematic. All Group 3 patients appeared LA-positive in undiluted plasma with Siemens dRVVT, as did 16/24 (67%) with HBM dRVVT but the fewer LA-positives in mixing tests better mapped to clear LA-positives with LD dRVVT. LD and Siemens dRVVTs exhibited 87% and 95% false-positivity for Group 6 whilst HBM dRVVT had none. Increasing the cut-off improved accuracy. Applying higher cut-offs improved accuracy in Group 4 patients.

Conclusion

HBM dRVVT exhibited improved specificity, mainly due to less interference by anticoagulation, but reduced sensitivity, compared to the other dRVVTs employed.

Keywords

Antiphospholipid antibodies antiphospholipid syndrome dilute Russell’s viper venom time false-positivity lupus anticoagulant reagent kits

Get full access to this article

View all access options for this article.

References

Miyakis

Lockshin

Atsumi

et al.

International consensus statement on an update of the classification criteria for definite antiphospholipid syndrome (APS). J Thromb Haemostasis 2006; 4: 295–306.

Moore

. Recent guidelines and recommendations for laboratory detection of lupus anticoagulants. Semin Thromb Hemostasis 2014; 40: 163–171.

Pengo

Tripodi

Reber

et al.

Subcommittee on Lupus Anticoagulant/Antiphospholipid Antibody of the Scientific and Standardisation Committee of the International Society on Thrombosis and Haemostasis. J Thromb Haemostasis 2009; 7: 1737–1740.

Keeling

Mackie

Moore

Greer

Greaves

. British Committee for Standards in Haematology. Guidelines on the investigation and management of antiphospholipid syndrome. Br J Haematol 2012; 157: 47–58.

Clinical and Laboratory Standards Institute. Laboratory Testing for the Lupus Anticoagulant; Approved Guideline. CLSI document H60-A, 2014. Wayne, PA: Clinical and Laboratory Standards Institute.

Lawrie

Mackie

Purdy

Machin

. The sensitivity and specificity of commercial reagents for the detection of lupus anticoagulant show marked differences in performance between photo-optical and mechanical coagulometers. Thromb Haemostasis 1999; 81: 758–762.

Jennings

Mackie

Arnout

Preston

. UK National External Quality Assessment Scheme for Blood Coagulation. Lupus anticoagulant testing using plasma spiked with monoclonal antibodies: performance in the UK NEQAS proficiency testing programme. J Thromb Haemostasis 2004; 2: 2178–2184.

McGlasson

Fritsma

. Comparison of six dilute Russell viper venom time lupus anticoagulant screen/confirm assay kits. Semin Thromb Hemostasis 2013; 39: 315–319.

Chantarangkul

Biguzzi

Asti

Palmucci

Tripodi

. Laboratory diagnostic outcome applying detection criteria recommended by the Scientific and Standardization Committee of the ISTH on Lupus Anticoagulant. Thromb Haemostasis 2013; 110: 46–52.

10.

Depreter

Devreese

. Dilute Russell’s viper venom time reagents in lupus anticoagulant testing: a well-considered choice. Clin Chem Lab Med 2017; 55: 91–101.

11.

Pennings

De Groot

Meijers

Huisman

Derksen

Urbanus

. Optimisation of lupus anticoagulant tests: should test samples always be mixed with normal plasma? Thromb Haemostasis 2014; 112: 736–742.

12.

Chandler

Torres

Rinder

Tormey

. Lupus anticoagulant testing and anticoagulation do not mix: quantitation of discrepant results and potential approaches to reduce false positives. Br J Haematol 2014; 167: 704–707.

13.

Krilis

Giannakopoulos

. Laboratory methods to detect antiphospholipid antibodies. Am Soc Hematol Hematol Educ Program 2014; 2014: 321–328.

14.

Martinuzzo

Barrera

D ’adamo

Otaso

Gimenez

Oyhamburu

. Frequent false-positive results of lupus anticoagulant tests in plasmas of patients receiving the new oral anticoagulants and enoxaparin. Int J Lab Hematol 2014; 36: 144–150.

15.

Ratzinger

Lang

Belik

et al.

Lupus-anticoagulant testing at NOAC trough levels. Thromb Haemostasis 2016; 116: 235–240.

16.

Gosselin

Grant

Adcock

. Comparison of the effect of the anti-Xa direct oral anticoagulants apixaban, edoxaban, and rivaroxaban on coagulation assays. Int J Lab Hematol 2016; 38: 505–513.

17.

Arachchillage

Mackie

Efthymiou

Isenberg

Machin

Cohen

. Interactions between rivaroxaban and antiphospholipid antibodies in thrombotic antiphospholipid syndrome. J Thromb Haemostasis 2015; 13: 1264–1273.

18.

Moore

. Combining Taipan snake venom time/Ecarin time screening with the mixing studies of conventional assays increases detection rates of lupus anticoagulants in orally anticoagulated patients. Thromb J 2007; 5: 125–125.

19.

van Os

de Laat

Kamphuisen

Meijers

de Groot

. Detection of lupus anticoagulant in the presence of rivaroxaban using Taipan snake venom time. J Thromb Haemostasis 2011; 9: 1657–1659.

20.

Moore

Brown

Bromidge

Drew

Ledford-Kraemer

. Lupus anticoagulant detection: out of control? Int J Lab Haematol 2013; 35: 128–136.

21.

Moore

Culhane

Daw

Noronha

Kumano

. Mixing test specific cut-off is more sensitive at detecting lupus anticoagulants than index of circulating anticoagulant. Thromb Res 2016; 139: 98–101.

22.

Gardiner

Mackie

Malia

et al.

The importance of locally derived reference ranges and standardized calculation of dilute Russell's viper venom time results in screening for lupus anticoagulant. Br J Haematol 2000; 111: 1230–1235.

23.

Devreese

KMJ

De Laat

. Mixing studies in lupus anticoagulant testing are required at least in some types of samples. J Thromb Haemostasis 2015; 13: 1475–1478.

24.

Depreter

Devreese

. Differences in lupus anticoagulant final conclusion through clotting time or Rosner index for mixing test interpretation. Clin Chem Lab Med 2016; 54: 1511–1516.

25.

Clinical and Laboratory Standards Institute. Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; Approved Guideline—Third Edition. CLSI document C28-A3, 2008. Wayne, PA: Clinical and Laboratory Standards Institute.

26.

Isert

Miesbach

Stoever

Lindhoff-Last

Linnemann

. Screening for lupus anticoagulants in patients treated with vitamin K antagonists. Int J Lab Hematol 2015; 37: 758–765.

27.

Pradella

Azzarini

Santarossa

et al.

Cooperation experience in a multicentre study to define the upper limits in a normal population for the diagnostic assessment of the functional lupus anticoagulant assays. Clin Chem Lab Med 2013; 51: 379–385.

28.

Reber

Meijer

. In ECAT veritas? Lupus 2012; 21: 722–724.

29.

Dembitzer

Ledford Kraemer

Meijer

Peerschke

. Lupus anticoagulant testing: performance and practices by North American clinical laboratories. Am J Clin Pathol 2010; 134: 764–773.

30.

Jacobsen

Wisløff

. False negative screening tests for lupus anticoagulants—an unrecognized problem? Thromb Res 1996; 82: 445–451.

31.

Giavarina

. Understanding Bland Altman analysis. Biochem Med 2015; 25: 141–151.

32.

Chighizola

Raschi

Banzato

Borghi

Pengo

Meroni

. The challenges of lupus anticoagulants. Expert Rev Hematol 2016; 9: 389–400.

33.

Jennings

Kitchen

Woods

Walker

. ISTH/SSC lupus anticoagulant testing guidelines: how far have these been adopted by laboratories? J Thromb Haemostasis 2011; 9: 2117–2119.

34.

Gerbutavicius

Fareed

Messmore

et al.

Reference intervals of the dilute tissue thromboplastin inhibition and dilute Russell’s viper venom tests revisited. Clin Appl Thromb Hemostasis 2002; 8: 115–124.

35.

Kumano

Ieko

Naito

Yoshida

Takahashi

Aoki

. Index of circulating anticoagulant cut-off value establishment in activated partial thromboplastin time mixing test for lupus anticoagulant diagnosis. J Thromb Haemostasis 2013; 110: 1919–1922.

36.

Tripodi

Chantarangkul

Cini

et al.

Variability of cut-off values for the detection of lupus anticoagulants. Results of an international multicenter multiplatform study. J Thromb Haemostasis. Epub ahead of print 17 March 2017. DOI: 10.1111/jth.13678.

37.

Sciascia

Breen

Hunt

. Rivaroxaban use in patients with antiphospholipid syndrome and previous venous thromboembolism. Blood Coagulation Fibrinolysis 2015; 26: 476–477.

38.

Cohen

Hunt

Efthymiou

et al.

RAPS trial investigators. Rivaroxaban versus warfarin to treat patients with thrombotic antiphospholipid syndrome, with or without systemic lupus erythematosus (RAPS): a randomised, controlled, open-label, phase 2/3, non-inferiority trial. Lancet Haematol 2016; 3: e426–e436.

39.

Pouplard

Vayne

Berthomet

et al.

The Taipan snake venom time can be used to detect lupus anticoagulant in patients treated by rivaroxaban. Int J Lab Hematol. Epub ahead of print 22 December 2016. DOI: 10.1111/ijlh.12611.

40.

Pengo

Biasiolo

Gresele

et al.

Participating Centres of Italian Federation of Thrombosis Centres (FCSA). Survey of lupus anticoagulant diagnosis by central evaluation of positive plasma samples. J Thromb Haemostasis 2007; 5: 925–930.