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DOI: 10.1055/s-0040-1721776
Inhibitor Index in the Clot Waveform Analysis-Based Mixing Test Differentiates among Hemophilia A without and with Inhibitors, and Lupus Anticoagulant
Funding This work was partly supported by a Grant-in-Aid for Scientific Research (KAKENHI) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) to K.N. (18K07885).Abstract
Background The mixing test is used to identify the pathway to follow-up testing and is also useful for the investigation of lupus anticoagulant (LA) positivity. “To completely correct” indicates clotting factor deficiency, while “to not correct” indicates the presence of a clotting factor inhibitor including LA. “Index of circulation anticoagulant” and/or “percent correction” is used to interpret the results of mixing studies, but it does not accurately differentiate factor inhibitors from LA.
Aim To precisely differentiate hemophilia A (HA), HA with inhibitor (HA-inh), and LA using the clot waveform analysis (CWA)-based mixing test.
Methods Plasma samples from HA, LA, and HA-inh including acquired HA were incubated with normal plasma in 9:1, 1:1, and 1:9 mix ratios. From activated partial thromboplastin time CWA at 0-minute (immediately) and 12-minute incubation, the ratios of CWA parameters at 12 minutes/0 minute (inhibitor index) were assessed.
Results The inhibitor index values of CWA parameters obtained using the mixing test in a 1:1 ratio demonstrated a significant difference between HA-inh and LA but could not differentiate LA from HA-inh completely. Plasmas used for the mixing tests in 9:1 and 1:9 ratios were able to fully distinguish between HA-inh (>0.5 BU/mL) and LA. These indices significantly correlated with inhibitor titer below 40 BU/mL (r > 0.90), possibly estimating FVIII inhibitor titer from the inhibitor index. Plasmas in HA and LA could be distinguished by mixing in a 1:1 ratio at 0 minute (immediately).
Conclusion The inhibitor index from CWA-based mixing tests with a 12-minute incubation could differentiate among HA, HA-inh, and LA quickly.
Note
An account of this work was presented at the XXVIII Congress of the International Society on Thrombosis and Haemostasis, 2020, Milano, Italy.
Authors' Contributions
N.S. performed the experiments, analyzed the data, and created the figures. K.O. analyzed and interpreted the data. Y.O., T.K., and M.E. performed the experiments and analyzed and interpreted the data. M.S. supervised this study. K.N. designed all the experiments, interpreted the data, created the figures, wrote and edited the manuscript, and approved the submission of the first version of the manuscript.
Publikationsverlauf
Eingereicht: 15. August 2020
Angenommen: 08. November 2020
Artikel online veröffentlicht:
07. Januar 2021
© 2021. Thieme. All rights reserved.
Georg Thieme Verlag KG
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References
- 1 Pengo V, Tripodi A, Reber G. et al; Subcommittee on Lupus Anticoagulant/Antiphospholipid Antibody of the Scientific and Standardisation Committee of the International Society on Thrombosis and Haemostasis. Update of the guidelines for lupus anticoagulant detection. J Thromb Haemost 2009; 7 (10) 1737-1740
- 2 Keeling D, Mackie I, Moore GW, Greer IA, Greaves M. British Committee for Standards in Haematology. Guidelines on the investigation and management of antiphospholipid syndrome. Br J Haematol 2012; 157 (01) 47-58
- 3 CLSI. Laboratory testing for the lupus anticoagulant; approved guideline, CLSI document H60-A. Wayne, PA: Clinical and Laboratory Standard Institute; 2014
- 4 Kershaw G. Performance and interpretation of mixing tests in coagulation. Methods Mol Biol 2017; 1646: 85-90
- 5 Kershaw G, Orellana D. Mixing tests: diagnostic aides in the investigation of prolonged prothrombin times and activated partial thromboplastin times. Semin Thromb Hemost 2013; 39 (03) 283-290
- 6 Rosner E, Pauzner R, Lusky A, Modan M, Many A. Detection and quantitative evaluation of lupus circulating anticoagulant activity. Thromb Haemost 1987; 57 (02) 144-147
- 7 Chang SH, Tillema V, Scherr D. A “percent correction” formula for evaluation of mixing studies. Am J Clin Pathol 2002; 117 (01) 62-73
- 8 Favaloro EJ, Bonar R, Duncan E. et al; RCPA QAP in Haematology Haemostasis Committee. Identification of factor inhibitors by diagnostic haemostasis laboratories: a large multi-centre evaluation. Thromb Haemost 2006; 96 (01) 73-78
- 9 Kershaw G. Detection and measurement of factor inhibitors. Methods Mol Biol 2017; 1646: 295-304
- 10 Sborov DW, Rodgers GM. Acquired hemophilia A: a current review of autoantibody disease. Clin Adv Hematol Oncol 2012; 10 (01) 19-27
- 11 Lossing TS, Kasper CK, Feinstein DI. Detection of factor VIII inhibitors with the partial thromboplastin time. Blood 1977; 49 (05) 793-797
- 12 Hay CR, Brown S, Collins PW, Keeling DM, Liesner R. The diagnosis and management of factor VIII and IX inhibitors: a guideline from the United Kingdom Haemophilia Centre Doctors Organisation. Br J Haematol 2006; 133 (06) 591-605
- 13 Favaloro EJ. Coagulation mixing studies: utility, algorithmic strategies and limitations for lupus anticoagulant testing or follow up of abnormal coagulation tests. Am J Hematol 2020; 95 (01) 117-128
- 14 Braun PJ, Givens TB, Stead AG. et al. Properties of optical data from activated partial thromboplastin time and prothrombin time assays. Thromb Haemost 1997; 78 (03) 1079-1087
- 15 Shima M, Thachil J, Nair SC, Srivastava A. Scientific and Standardization Committee. Towards standardization of clot waveform analysis and recommendations for its clinical applications. J Thromb Haemost 2013; 11 (07) 1417-1420
- 16 Shima M, Matsumoto T, Fukuda K. et al. The utility of activated partial thromboplastin time (aPTT) clot waveform analysis in the investigation of hemophilia A patients with very low levels of factor VIII activity (FVIII:C). Thromb Haemost 2002; 87 (03) 436-441
- 17 Matsumoto T, Nogami K, Tabuchi Y. et al. Clot waveform analysis using CS-2000i™ distinguishes between very low and absent levels of factor VIII activity in patients with severe haemophilia A. Haemophilia 2017; 23 (05) e427-e435
- 18 Haku J, Nogami K, Matsumoto T, Ogiwara K, Shima M. Optimal monitoring of bypass therapy in hemophilia A patients with inhibitors by the use of clot waveform analysis. J Thromb Haemost 2014; 12 (03) 355-362
- 19 Ochi S, Takeyama M, Shima M, Nogami K. Plasma-derived factors VIIa and X mixtures (Byclot®) significantly improve impairment of coagulant potential ex vivo in plasmas from acquired hemophilia A patients. Int J Hematol 2020; 111 (06) 779-785
- 20 Matsumoto T, Nogami K, Shima M. A combined approach using global coagulation assays quickly differentiates coagulation disorders with prolonged aPTT and low levels of FVIII activity. Int J Hematol 2017; 105 (02) 174-183
- 21 Shimonishi N, Ogiwara K, Oda Y. et al. A novel assessment of factor VIII activity by template matching utilizing weighted average parameters from comprehensive clot waveform analysis. Thromb Haemost 2021; 121 (02) 164-173
- 22 Moore GW, Culhane AP, Daw CR, Noronha CP, Kumano O. Mixing test specific cut-off is more sensitive at detecting lupus anticoagulants than index of circulating anticoagulant. Thromb Res 2016; 139: 98-101
- 23 Kumano O, Moore GW. Lupus anticoagulant mixing tests for multiple reagents are more sensitive if interpreted with a mixing test-specific cut-off than index of circulating anticoagulant. Res Pract Thromb Haemost 2017; 2 (01) 105-113
- 24 Kumano O, Moore GW. Ruling out lupus anticoagulants with mixing test-specific cutoff assessment and the index of circulating anticoagulant. Res Pract Thromb Haemost 2019; 3 (04) 695-703
- 25 Moore GW. Recent guidelines and recommendations for laboratory detection of lupus anticoagulants. Semin Thromb Hemost 2014; 40 (02) 163-171
- 26 Meroni PL, Borghi MO, Raschi E, Tedesco F. Pathogenesis of antiphospholipid syndrome: understanding the antibodies. Nat Rev Rheumatol 2011; 7 (06) 330-339
- 27 Kasper CK, Aledort L, Aronson D. et al. Proceedings: a more uniform measurement of factor VIII inhibitors. Thromb Diath Haemorrh 1975; 34 (02) 612
- 28 Kumano O, Ieko M, Naito S, Yoshida M, Takahashi N. APTT reagent with ellagic acid as activator shows adequate lupus anticoagulant sensitivity in comparison to silica-based reagent. J Thromb Haemost 2012; 10 (11) 2338-2343
- 29 Dumoulin EN, Fiers L, Devreese KM. Investigation of sensitivity for coagulation factor deficiency in APTT and PT: how to perform it?. Clin Chem Lab Med 2016; 54 (05) e169-e172
- 30 Martinuzzo M, Barrera L, Rodriguez M, D'Adamo MA, López MS, Otaso JC. Do PT and APTT sensitivities to factors' deficiencies calculated by the H47-A2 2008 CLSI guideline reflect the deficiencies found in plasmas from patients?. Int J Lab Hematol 2015; 37 (06) 853-860
- 31 Pengo V, Bison E, Banzato A, Zoppellaro G, Jose SP, Denas G. Lupus anticoagulant testing: diluted Russell viper venom time (dRVVT). Methods Mol Biol 2017; 1646: 169-176