Lupus anticoagulant mixing tests for multiple reagents are more sensitive if interpreted with a mixing test‐specific cut‐off than index of circulating anticoagulant
Open accessAbstract
Unlabelled Box •Various approaches are available for interpreting lupus anticoagulant mixing tests. •Mixing test specific cut‐off (MTC) was compared with index of circulating anticoagulant (ICA). •MTC was superior to ICA in detecting inhibition by lupus anticoagulant (LA) in multiple reagents. •Detection rates of LA can be improved by the method to interpret the results. Background Lupus anticoagulant (LA) is classified in the antibody family that is recognized as antiphospholipid antibodies. Guidelines for LA detection recommend mixing test interpretation with either a mixing test specific cut‐off (MTC) or index of circulating anticoagulant (ICA). We previously evidenced that MTC was superior to ICA in detecting the in vitro inhibition of LA with a single dilute APTT (activated partial thromboplastin time) and dRVVT (diluted Russell's viper venom time) pairing. Objectives The objective in the present study was to compare the LA diagnostic effectiveness of MTC and ICA by multiple APTT and dRVVT reagents. Methods One hundred‐five samples from non‐anticoagulated patients positive for LA in the dilute APTT (dAPTT) and dRVVT reagent pairing employed for diagnostic examination were performed by undiluted and in a 1:1 mix with normal pooled plasma with four additional APTT reagents and another dRVVT reagent (dRVVT B). Results Frequencies of MTC and ICA positivity were determined from samples LA positive in undiluted plasma. MTC positivity in mixing test were 63%, 77%, 80%, 84%, 46%, 81%, and 72% in 4 APTT, dAPTT and 2 dRVVT, respectively. ICA positivity were 47%, 67%, 58%, 54%, 42%, 47%, and 29%, respectively. There were no samples of ICA‐positive/MTC‐negative with any reagent. Conclusions The data indicate that MTC is superior to ICA for LA detection in mixing tests in multiple reagents and reagent types. Although mixing tests may make weak LA samples appear negative, the efficacy of LA detection can be improved by the method to interpret the results.
AI snapshot
Written from the paper’s full text.
TL;DR
In 105 non-anticoagulated patients already positive for lupus anticoagulant (LA), interpreting 1:1 mixing tests with a mixing test-specific cut-off (MTC) detected more LA than the index of circulating anticoagulant (ICA) across multiple APTT and dRVVT reagents. No sample was ICA-positive/MTC-negative, supporting MTC as the more sensitive mixing-test interpretation.
Key findings
- MTC positivity in mixing tests was 63%, 77%, 80%, 84%, 46%, 81%, and 72% across the four APTT reagents, dAPTT, and two dRVVT reagents, respectively; ICA positivity was 47%, 67%, 58%, 54%, 42%, 47%, and 29%, respectively.
- There were no samples that were ICA-positive/MTC-negative with any reagent.
- In undiluted plasma, LA positivity ranged from 33% to 63% for APTT-based assays and 62% to 70% for dRVVT assays.
- Mix ratio and ICA values were significantly higher in samples positive by both MTC and ICA than in MTC-only or both-negative groups in all reagents (P < .01).
Methods
- Plasma samples from 105 non-anticoagulated patients who were LA-positive on routine diagnostic testing: 44 with definite APS, 12 with SLE and persistent LA, 9 with SLE and LA-positive at testing, and 40 undergoing diagnostic testing for aPL/APS.
- Samples were tested undiluted and in 1:1 mixes with normal pooled plasma using four additional APTT reagents and another dRVVT reagent (dRVVT B), alongside the routine dAPTT and dRVVT reagents.
- Cut-offs were derived from 50 normal samples; comparisons used Wilcoxon signed rank tests with P < .01 considered significant, and reference intervals were calculated as mean ± 2 SD.
Limitations
- The paper states that mixing tests were assessed in the “ideal” situation of otherwise uncompromised samples, and that further work is planned in other situations including anticoagulant therapy and factor deficiencies, with ROC curve analysis for cut-off generation.
- The paper notes that mixing tests introduce a dilution factor and may make weak LA samples appear negative.
- Evident from the text: because only LA-positive samples were studied, the findings address sensitivity of MTC versus ICA but do not establish specificity or performance in LA-negative or unselected populations.
Model-written summary. Check it against the paper before relying on it.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
10 of 31 cited sources checked · 7 supported · 0 flagged · accuracy not yet measured
- Premisecitation
Mixing tests may make weak LA samples appear negative due to a dilution factor.
“mixing plasma with NPP shows a dilution factor that can make weaker LA samples appear negative in the mixing test despite clear positivity in the screening and confirmation results on undiluted plasma”
- Also stated in: § Conclusions · hedged
“Although mixing tests may make weak LA samples appear negative”
Quote in the paperSubstantial claim1 of 3 sources checked- cites [7] 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: 47– 58. 22313321 source
- cites [8] Ledford‐Kraemer MR, Moore GW, Bottenus R, Danniele C, de Groot PG, Exner T. Anticoagulant Approved Guideline. 1st ed CLSI Document H60. Wayne, PA: Clinical and Laboratory Standards Institute; 2014.
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
- cites [13] Devreese KM. Interpretation of normal plasma mixing studies in the laboratory diagnosis of lupus anticoagulants. Thromb Res. 2007; 119: 369– 76. 16704874 source
- Also stated in: § Conclusions · hedged
- Premisecitation
In situations where alternative or co-existing coagulation abnormalities are present, mixing tests can improve specificity of LA testing.
“However, in situations where alternative or co‐existing coagulation abnormalities are present, mixing tests can improve specificity of LA testing.34, 35, 36, 37”
Quote in the paperNot a substantial claim1 of 3 sources checked- cites [34] Devreese KMJ, Laat BD. Mixing studies in lupus anticoagulant testing are required at least in some type of samples. J Thromb Haemost. 2015; 13: 1– 4. 26017873 source
- cites [35] Martinuzzo ME, Barrera LH, Dadamo 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 Hematol. 2014; 36: 144– 50. 24034808 source
- cites [36] 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. 25008787 source
- Premisecitation
Potency alone does not necessarily correlate with clinical significance and thrombotic risk.
“On the other hand, a number of studies have indicated populations of clinically significant LA despite the negative results in the mixing tests14, 32, 36 and the potency alone does not necessarily correlate with clinical significance and thrombotic risk.37, 39”
Quote in the paperNot a substantial claim2 of 2 sources checked- cites [37] Devreese KMJ. No more mixing tests required for integrated assay systems in the laboratory diagnosis of lupus anticoagulant? J Thromb Haemost. 2010; 8: 1120– 2. 20149080 source
- cites [39] Devreese K, Peerlinck K, Hoylaerts MF. Thrombotic risk assessment in the antiphospholipid syndrome requires more than the quantification of lupus anticoagulants. Blood. 2010; 115: 870– 8. 19965629 source
- Findingown data
Positivity by both MTC and ICA was higher with FSL than all other reagents in the study.
“In addition, positivity by both MTC and ICA was higher with FSL than all other reagents in the study.”
Quote in the paperSubstantial claim- Rests on an assumption: The ICA cut-off values established with the routinely employed LA reagents are applicable to the additional APTT and dRVVT reagents.
- Findingown data
Mix ratio values of MTC only positive samples were significantly higher than those negative in both MTC and ICA in all reagents.
“In addition, the values of MTC only positive were significantly higher than those of negative in both MTC and ICA in all reagents.”
Quote in the paperSubstantial claim
- Findingown data
ICA values of MTC only positive samples were significantly higher than those negative in both MTC and ICA in all reagents.
“Although the values in two groups such as MTC only positive and negative in both MTC and ICA were under the cut‐off value, the values in MTC only positive group were significantly higher than those of negative both MTC and ICA group in all reagents.”
Quote in the paperSubstantial claim- Rests on an assumption: The ICA cut-off values established with the routinely employed LA reagents are applicable to the additional APTT and dRVVT reagents.
- Findingown data
No samples were positive by ICA alone (ICA-positive/MTC-negative) in any reagent.
“There were no samples of ICA‐positive/MTC‐negative with any reagent.”
- Also stated in: § DISCUSSION · plain
“there were no positive samples for inhibition by ICA alone in any reagent.”
Quote in the paperSubstantial claim- Rests on an assumption: The ICA cut-off values established with the routinely employed LA reagents are applicable to the additional APTT and dRVVT reagents.
- Also stated in: § DISCUSSION · plain
- Findingown data
In LA-positive samples, MTC positivity in mixing tests was higher than ICA positivity across multiple APTT and dRVVT reagents.
“MTC was superior to ICA in detecting inhibition by lupus anticoagulant (LA) in multiple reagents.”
- Also stated in: § Comparison of positivity in MTC and ICA · plain
“Frequency of MTC‐positivity was higher for ICA in all reagents.”
- Also stated in: § DISCUSSION · boosted
“Without exception, MTC showed higher sensitivity than ICA in detection of known LA with every reagent.”
Quote in the paperSubstantial claim- Rests on an assumption: The ICA cut-off values established with the routinely employed LA reagents are applicable to the additional APTT and dRVVT reagents.
- Also stated in: § Comparison of positivity in MTC and ICA · plain
- Findingown data
Mean and median screen ratios in undiluted plasma were higher in samples positive by both MTC and ICA than in those positive by MTC alone in all reagents.
“Mean and median screen ratios were higher in samples positive by MTC and ICA than those positive by MTC alone in all reagents.”
Quote in the paperNot a substantial claim - Findingown data
Mix ratio values of samples positive by both MTC and ICA were significantly higher than those of MTC only and negative in both MTC and ICA in all reagents.
“The mix ratio values of samples positive in mixing test by both MTC and ICA were significantly higher than those of MTC only and negative in both MTC and ICA in all reagents.”
Quote in the paperSubstantial claim - Findingown data
ICA positivity frequencies were 47%, 67%, 58%, 54%, 42%, 47%, and 29%, respectively.
“ICA positivity were 47%, 67%, 58%, 54%, 42%, 47%, and 29%, respectively.”
Quote in the paperSubstantial claim- Rests on an assumption: The ICA cut-off values established with the routinely employed LA reagents are applicable to the additional APTT and dRVVT reagents.
- Findingown data
MTC positivity frequencies in mixing tests were 63%, 77%, 80%, 84%, 46%, 81%, and 72% in 4 APTT, dAPTT and 2 dRVVT, respectively.
“MTC positivity in mixing test were 63%, 77%, 80%, 84%, 46%, 81%, and 72% in 4 APTT, dAPTT and 2 dRVVT, respectively.”
Quote in the paperSubstantial claim - Findingown data
ICA values of samples positive by both MTC and ICA were significantly higher than those of MTC only and negative in both MTC and ICA in all reagents.
“The ICA values of samples positive in mixing test by both MTC and ICA were significantly higher than those of MTC only and negative in both MTC and ICA in all reagents.”
Quote in the paperSubstantial claim- Rests on an assumption: The ICA cut-off values established with the routinely employed LA reagents are applicable to the additional APTT and dRVVT reagents.
- Interpretationown data
MTC is superior to ICA for LA detection in mixing tests in multiple reagents and reagent types.
“The data indicate that MTC is superior to ICA for LA detection in mixing tests in multiple reagents and reagent types.”
- Also stated in: § DISCUSSION · hedged
“our data indicate that MTC had higher sensitivity than ICA for detecting LA in multiple reagents.”
Quote in the paperSubstantial claim- Rests on an assumption: The ICA cut-off values established with the routinely employed LA reagents are applicable to the additional APTT and dRVVT reagents.
- Also stated in: § DISCUSSION · hedged
- Interpretationown data
Ellagic acid-based APTT reagents are not necessarily less effective than others in detecting LA.
“These data provide further evidence, in a large clinically appropriate population, that ellagic acid‐based APTT reagents are not necessarily less effective than others in detecting LA.”
Quote in the paperSubstantial claim - Premisecitation
Sensitivity and specificity of different APTT and dRVVT reagents to LA varies, predominantly due to differences in phospholipid composition and concentration.
“Sensitivity and specificity of different APTT and dRVVT reagents to LA varies, predominantly due to differences in phospholipid composition and concentration.10, 11, 12”
Quote in the paperSubstantial claim0 of 3 sources checked- cites [10] Brandt JT, Triplett DA, Musgrave K, Orr C. The sensitivity of different coagulation reagents to the presence of lupus anticoagulants. Arch Pathol Lab Med. 1987; 111: 120– 4. 3101649 source
- cites [11] Denis‐Magdelaine A, Flahault A, Verdy E. Sensitivity of sixteen APTT reagents for the presence of lupus anticoagulants. Haemostasis. 1995; 25: 98– 105. 7607585 source
- cites [12] 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: 2338– 43. 22909048 source
- Premisecitation
In a previous study, MTC had higher sensitivity than ICA for detecting in vitro inhibition of LA with a single dAPTT and dRVVT reagent pairing.
“We previously evidenced that MTC was superior to ICA in detecting the in vitro inhibition of LA with a single dilute APTT (activated partial thromboplastin time) and dRVVT (diluted Russell's viper venom time) pairing.”
- Also stated in: § DISCUSSION · plain
“as previously described with single APTT and dRVVT reagent pairings.9, 30”
Quote in the paperSubstantial claim0 of 2 sources checked- cites [9] 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. 26916303 source
- cites [30] Depreter B, Devreese KM. Differences in lupus anticoagulant final conclusion through clotting time or Rosner index for mixing test interpretation. Clin Chem Lab Med. 2016; 54: 1511– 9. 26894579 source
- Also stated in: § DISCUSSION · plain
- Interpretationcitation
Manifestation in mixing tests seems to be a function of more than just antibody potency; other possible contributory factors include epitope specificity, antibody avidity, and reagent composition.
“Manifestation in mixing tests seems to be a function of more than just potency, other possible contributory factors include epitope specificity, antibody avidity, and reagent composition.9”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [9] 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. 26916303 source
- Interpretationreasoning
Maximizing diagnostic capability of mixing tests for when they are needed, by applying the most sensitive interpretive index, improves the efficacy of LA detection.
“Although mixing tests introduce a dilution factor and may make weak LA samples appear negative, maximizing diagnostic capability of mixing tests for when they are needed, by applying the most sensitive interpretive index, improves the efficacy of LA detection.”
Quote in the paperSubstantial claim- Rests on an assumption: The ICA cut-off values established with the routinely employed LA reagents are applicable to the additional APTT and dRVVT reagents.
- Interpretationreasoning
The LA-sensitivity of a given reagent is a crucial contributor to the efficacy of mixing tests performed with it.
“The LA‐sensitivity of a given reagent is a crucial contributor to the efficacy of mixing tests performed with it, and interpretation via MTC appears to enhance mixing test sensitivity.”
Quote in the paperSubstantial claim - Premisecitation
The mixing of weak LA samples may introduce loss of detectable LA activity and false-negativity give rise to inaccurate interpretation if the mixing test result is employed as a decision point for subsequent confirmatory test performance.
“The mixing of weak LA samples may introduce loss of detectable LA activity and the false‐negativity give rise to inaccurate interpretation if the mixing test result is employed as a decision point for subsequent confirmatory test performance.14, 31, 32, 33”
Quote in the paperSubstantial claim0 of 3 sources checked- cites [14] 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. 16263154 source
- cites [31] Hong SK, Hwang SM, Kim JE, Kim HK. Clinical significance of the mixing test in laboratory diagnoses of lupus anticoagulant: the fate of the mixing test in integrated lupus anticoagulant test systems. Blood Coagul Fibrinolysis. 2012; 23: 739– 44. 22964767 source
- cites [32] Thom J, Ivey L, Eikelboom J. Normal plasma mixing studies in the laboratory diagnosis of lupus anticoagulant. J Thromb Haemost. 2003; 1: 2689– 91. 14675112 source
- Interpretationown data
Stronger antibodies (higher screen ratio in undiluted plasma) are more likely to generate elevations of both MTC and ICA.
“Using screen ratio in undiluted plasma as indicator of antibody potency, the data suggest that stronger antibodies are more likely to generate elevations of both MTC and ICA.”
Quote in the paperNot a substantial claim
- Classified as probably not a substantial claim. 10
- May combine two claims. 4
- The quote only partly states this claim. 1
Classified as probably not a substantial claim.
PremisecitationGuidelines recommend that mixing tests are interpreted with either MTC or ICA.
“Guidelines recommend mixing tests are interpreted with either a mixing test specific cut‐off (MTC) or the index of circulating anticoagulant (ICA).”
Quote in the paperSubstantial claim2 of 3 sources checked- cites [6] 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: 1737– 40. 19624461 source
- cites [7] 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: 47– 58. 22313321 source
The Lupus anticoagulant testing section says mixing tests may be used to detect an inhibitor, and the Recommendation section describes interpreting an abnormal screening result from a mixture. Neither passage recommends interpreting mixing tests with an MTC or ICA; the paper’s discussion of cut-offs concerns LA tests generally, not those two methods specifically. Page numbers are unavailable in our copy. (LLM, conf=0.84)
Experimental — accuracy not yet measured - cites [8] Ledford‐Kraemer MR, Moore GW, Bottenus R, Danniele C, de Groot PG, Exner T. Anticoagulant Approved Guideline. 1st ed CLSI Document H60. Wayne, PA: Clinical and Laboratory Standards Institute; 2014.
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
Classified as probably not a substantial claim. May combine two claims.
PremisecitationThere is no single test to detect all LA, and guidelines recommend performance of two different tests (dRVVT and LA-sensitive APTT).
“There is no single test to detect all LA1, 6, 7, 8 and guidelines recommend performance of two different tests that represent different assay principles, the diluted Russell's viper venom time (dRVVT) and a LA‐sensitive activated partial thromboplastin time (APTT).6, 7, 8”
Quote in the paperSubstantial claim2 of 3 sources checked- cites [1] Meroni PL, Borghi O, Raschi E, Tedesco F. Pathogenesis of antiphospholipid syndrome: understanding the antibodies. Nat Rev Rheumatol. 2011; 7: 330– 9. 21556027 source
- cites [6] 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: 1737– 40. 19624461 source
- cites [7] 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: 47– 58. 22313321 source
Probably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationIntegrated testing is sufficient to detect LA without performing the mixing test in samples with no other causes of clotting time prolongation and LA confirmatory test results within reference intervals.
“For samples from patients with no other causes of clotting time prolongation and LA assay confirmatory test results within reference intervals, the increasingly popular paradigm of integrated testing is sufficient to detect the presence of LA without performing the mixing test at all.6, 7, 8, 9, 26, 27, 28, 31, 33, 34”
Quote in the paperNot a substantial claim2 of 3 sources checked- cites [6] 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: 1737– 40. 19624461 source
The Integrated tests section says these tests combine screening and confirmation and, in principle, do not require a mixing test. It does not establish that they are sufficient specifically when other causes of prolonged clotting time are absent and confirmatory results fall within reference intervals. Its statement is also more qualified than the citing sentence’s assertion of sufficiency. (LLM, conf=0.83)
Experimental — accuracy not yet measured - cites [7] 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: 47– 58. 22313321 source
The Lupus anticoagulant testing section says that, when no other cause of prolonged clotting time is present, samples may be considered LA-positive based on positive screening and confirmatory tests on undiluted plasma, despite a negative mixing test. It does not specify that the confirmatory test result must fall within its reference interval, and it recommends confirmation with a fresh sample whenever possible. Page numbers are unavailable in our copy. (LLM, conf=0.84)
Experimental — accuracy not yet measured - cites [8] Ledford‐Kraemer MR, Moore GW, Bottenus R, Danniele C, de Groot PG, Exner T. Anticoagulant Approved Guideline. 1st ed CLSI Document H60. Wayne, PA: Clinical and Laboratory Standards Institute; 2014.
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
May combine two claims.
Findingown dataThe percentage of LA detected in the FSL screening test was 57%, higher than SP (38%) and SLA (33%), and similar to Cephen and PTT silica-based reagents.
“In the present study, performed on a much larger cohort of LA‐containing plasmas, the percentage of LA detected in the FSL screening test was 57%, much higher than the 38% and 33%, respectively, of SP and SLA, and similar to that of Cephen and PTT silica‐based reagents.”
Quote in the paperSubstantial claimMay combine two claims.
FindingcitationScreen and mix ratios and ICA values were similar between the two dRVVT reagents.
“Screen and mix ratios and ICA values were similar between the two dRVVT reagents, possibly due, at least in part, to normalizing the data reducing between reagent differences.28, 29”
Quote in the paperNot a substantial claim0 of 2 sources checked- cites [28] McGlasson DL, Fritsma GA. Comparison of six dilute Russell viper venom time lupus anticoagulant screen/confirm assay kits. Semin Thromb Hemost. 2013; 39: 315– 9. 23468380 source
- cites [29] Jennings I, Greaves M, Mackie IJ, Kitchen S, Woods TA, Preston FE; UK National External Quality Assessment Scheme for Blood Coagulation. Lupus anticoagulant testing: improvements in performance in a UK NEQAS proficiency testing exercise after dissemination of national guidelines on laboratory methods. Br J Haematol. 2002; 119: 364– 9. 12406070 source
Probably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductown dataLA2 confirmation reagent confirmed LA activity in undiluted plasma for all samples with elevated LA1 screening results.
“Similarly, LA2 confirmation reagent (Siemens Healthineers) confirmed LA activity in undiluted plasma for all samples with elevated LA1 screening results.”
Quote in the paperNot a substantial claimThe quote only partly states this claim.
Interpretationown dataDetection rates of LA can be improved by using MTC rather than ICA to interpret mixing test results.
“Detection rates of LA can be improved by the method to interpret the results.”
- Also stated in: § Conclusions · plain
“the efficacy of LA detection can be improved by the method to interpret the results.”
Quote in the paperSubstantial claim- Rests on an assumption: The ICA cut-off values established with the routinely employed LA reagents are applicable to the additional APTT and dRVVT reagents.
- Also stated in: § Conclusions · plain
Probably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductcitationAll samples with elevated APTT screen ratios achieved significant correction by the Actin FS ratio to confirm LA activity in undiluted plasma.
“All samples with elevated APTT screen ratios achieved significant correction by the Actin FS ratio to confirm LA activity in undiluted plasma.”
Quote in the paperNot a substantial claimClassified as probably not a substantial claim.
Study design and conductcitationCut-off values for screen, confirm, and mixing test in each additional reagent were determined from upper limits of the distribution of 50 normal samples in each reagent.
“Cut‐off values for screen, confirm, and mixing test in each additional reagent were determined from upper limits of the distribution of 50 normal samples in each reagent, and for ICA with the routinely employed LA reagents, ICA cut‐off values were previously established and used in each additional reagent.9, 21, 22, 23”
Quote in the paperSubstantial claim0 of 3 sources checked- cites [9] 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. 26916303 source
- cites [21] Kumano O, Ieko M, Naito S, Yoshida M, Takahashi N, Aoki T. Index of circulating anticoagulant cut‐off value establishment in activated partial thromboplastin time mixing test for lupus anticoagulant diagnosis. J Thromb Haemost. 2013; 11: 1919– 22. 23819748 source
- cites [22] Kumano O, Ieko M, Naito S, 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. 24935674 source
Classified as probably not a substantial claim.
Study design and conductnone givenFactor deficiencies and undisclosed anticoagulation were excluded by performing coagulation screening tests prior to the LA assays.
“Factor deficiencies and undisclosed anticoagulation were excluded by performing coagulation screening tests prior to the LA assays.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductnone given53.3% of the cohort had established persistent LA.
“We performed the LA assays in a clinically select and appropriate cohort, 53.3% of whom had established persistent LA.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductnone givenThe population distributions for the normal donor samples were confirmed as Gaussian.
“The population distributions for the normal donor samples were confirmed as Gaussian.”
Quote in the paperNot a substantial claimClassified as probably not a substantial claim.
Study design and conductreasoningThe study specifically assessed mixing tests in the 'ideal' situation of otherwise uncompromised samples.
“Thus, it is valuable to maximise diagnostic efficacy of mixing tests, so for the present study, we specifically assessed mixing tests in the “ideal” situation of otherwise uncompromised samples.”
Quote in the paperSubstantial claimMay combine two claims.
InterpretationreasoningWhilst integrated testing can be diagnostically accurate and logistically attractive, there are situations where additionally undertaking mixing tests achieves accurate and more confident diagnoses.
“Whilst integrated testing can be diagnostically accurate and logistically attractive, there are situations where additionally undertaking mixing tests achieves accurate and more confident diagnoses.”
Quote in the paperSubstantial claim
The paper relies on this without support.
The ICA cut-off values established with the routinely employed LA reagents are applicable to the additional APTT and dRVVT reagents.
“for ICA with the routinely employed LA reagents, ICA cut‐off values were previously established and used in each additional reagent.9, 21, 22, 23”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
ICA has a high specificity to factor deficiency, whereas the mixing test ratio has a high sensitivity to inhibitors.
“For instance, ICA has a high specificity to factor deficiency, whereas the mixing test ratio has a high sensitivity to inhibitors [6,7], although both indexes are recommended for LA diagnosis [8,9].”
From Performance characteristics of 5 numerical indexes in mixing test interpretation under coexistence of lupus anticoagulant and coagulation factor deficiency (2023) · cites this paper as [6]
The Discussion reports that a mixing test-specific cut-off (MTC) was more sensitive than ICA for detecting lupus anticoagulant, supporting the sensitivity comparison for this inhibitor. The Materials and Methods section says factor deficiencies were excluded before testing; the paper therefore does not establish that ICA has high specificity to factor deficiency. (LLM, conf=0.88)
Experimental — accuracy not yet measuredPrevious studies have improved the sensitivity and specificity by establishing suitable cutoff values.
“Previous studies have improved the sensitivity and specificity by establishing suitable cutoff values [1,6].”
From Performance characteristics of 5 numerical indexes in mixing test interpretation under coexistence of lupus anticoagulant and coagulation factor deficiency (2023) · cites this paper as [6]
The Discussion (page unavailable) reports that a mixing-test-specific cut-off improved sensitivity compared with the index of circulating anticoagulant. It says mixing tests can improve specificity in certain circumstances, but does not show that establishing the cut-off improved specificity. (LLM, conf=0.85)
Experimental — accuracy not yet measuredThe mixing test normalized ratio is most sensitive to inhibitors.
“The mixing test normalized ratio showed non-correction in all LA samples, indicating that this index is most sensitive to inhibitors”
From Performance characteristics of 5 numerical indexes in mixing test interpretation under coexistence of lupus anticoagulant and coagulation factor deficiency (2023) · cites this paper as [6]
The Discussion (page not provided) says that the mixing ratio interpreted with a mixing-test-specific cut-off was more sensitive than the index of circulating anticoagulant for detecting lupus anticoagulant inhibition. It does not establish that the ratio is most sensitive to inhibitors generally; the Results (page not provided) also show that not all lupus-anticoagulant-positive samples were positive in mixing tests. (LLM, conf=0.83)
Experimental — accuracy not yet measuredAlthough MTC provides greater sensitivity for the detection of LA, especially when weak LA is present, it has a greater tendency to produce false-positive results than ICA.
“Although MTC provides greater sensitivity for the detection of LA, especially when weak LA is present, it has a greater tendency to produce false-positive results than ICA [16,18,19].”
From Comparison of different algorithms for lupus anticoagulant detection: a single-center experience (2024) · cites this paper as [18]
The Results and Discussion report higher MTC sensitivity than ICA for detecting LA, including cases that mixing-test dilution could otherwise cause to appear negative. But the reported comparison uses samples positive for LA in undiluted plasma; it does not establish that MTC produces more false-positive results than ICA. No page numbers are available in our copy. (LLM, conf=0.88)
Experimental — accuracy not yet measuredPerformance characteristics in each index have been well examined in samples with a single cause of APTT prolongation, but those with 2 causes remain largely unknown.
“Although performance characteristics in each index have been well examined in samples with a single cause of APTT prolongation [1,[5], [6], [7]], those with 2 causes (eg, factor VIII [FVIII] deficiency and LA) remain largely unknown.”
MTC and ICA can be calculated and employed as interpretation indices for mixing tests.
“In addition, the mixing test–specific cutoff (MTC) and index of circulating anticoagulant (ICA) can be calculated and employed as interpretation indices for the mixing tests [[15], [16], [17], [18], [19]].”
From Comparison of different algorithms for lupus anticoagulant detection: a single-center experience (2024) · cites this paper as [18]
Several studies have recommended the use of MTC over ICA as it offers greater sensitivity in detecting in vitro inhibition of LA.
“For the interpretation of the LA mixing test result, several studies have recommended the use of MTC over ICA as it offers greater sensitivity in detecting in vitro inhibition of LA [[17], [18], [19]].”
From Comparison of different algorithms for lupus anticoagulant detection: a single-center experience (2024) · cites this paper as [18]
ICA has recently been demonstrated to be less sensitive to LA than a mixing-test-specific cut-off expressed as a normalized ratio.
“Nevertheless, ICA, originally developed for LA detection and later exported to the interpretation of other mixing tests in hemostasis [56], has recently been demonstrated to be less sensitive to LA than a mixing-test-specific cut-off expressed as a normalized ratio [57,58,59].”
From Lupus Anticoagulant Detection under the Magnifying Glass (2023) · cites this paper as [59]
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Self-citationScreening for lupus anticoagulant: improving the performance of the lupus-sensitive PTT-LA
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Flagged first, then by citations
2 of the 44 references couldn’t be matched to a known paper.
Cited by
Sorted by citations
Favaloro, E. J. (2020) · American Journal of Hematology · doi:10.1002/ajh.25669 · Cited by 53 · see citing works
Activated Partial Thromboplastin Time and Prothrombin Time Mixing Studies: Current State of the Art
Adcock, D. M., Moore, G. W., Montalvão, S. A. D. L., Kershaw, G. W., & Gosselin, R. C. (2022) · Seminars in Thrombosis and Hemostasis · doi:10.1055/s-0042-1756196 · Cited by 29 · see citing works
Self-citationTesting for Lupus Anticoagulants
Moore, G. W. (2022) · Seminars in Thrombosis and Hemostasis · doi:10.1055/s-0042-1744363 · Cited by 30 · see citing works
Self-citationKumano, O., & Moore, G. W. (2019) · Research and Practice in Thrombosis and Haemostasis · doi:10.1002/rth2.12245 · Cited by 24 · see citing works
Self-citationMixing studies for lupus anticoagulant: mostly no, sometimes yes
Moore, G. W. (2020) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm-2019-1248 · Cited by 19 · see citing works
“A variety of formulae have been proposed and employed for interpreting mixing tests and current guidelines recommend either the index of circulating anticoagulant (ICA) calculation, or a mixing test-specific cut-off (MTC) derived from the upper limit of a population distribution [11, 17].”
BackgroundInfluentialSelf-citationMoore, G. W., & Kumano, O. (2020) · Journal of Thrombosis and Haemostasis · doi:10.1111/jth.14669 · Cited by 18 · see citing works
Self-citationEvaluation of an automated algorithm for interpretation of lupus anticoagulant testing
Florin, L., Desloovere, M., & Devreese, K. M. J. (2019) · International Journal of Laboratory Hematology · doi:10.1111/ijlh.13001 · Cited by 12 · see citing works
Variability among commercial batches of normal pooled plasma in lupus anticoagulant testing
Cabo, J., Morimont, L., Baudar, J., Guldenpfennig, M., Jacqmin, H., Soleimani, R., Lecompte, T., Douxfils, J., & Mullier, F. (2022) · International Journal of Laboratory Hematology · doi:10.1111/ijlh.13977 · Cited by 12 · see citing works
“Based on recent publications, the current ISTH guideline recommends the use of a cut-off specific for mixing test ratio and against the use of ICA, because the latter has been shown to be less sensitive to LA inhibition.(3,15,16,20,21) Although ICA tends to be abandoned in favor of TABLE 2 Comparison of CTs of two sou…”
MethodKumano, O., Amiral, J., Dunois-Lardé, C., Peyrafitte, M., & Moore, G. W. (2018) · International Journal of Laboratory Hematology · doi:10.1111/ijlh.12921 · Cited by 11 · see citing works
Self-citationLupus Anticoagulant Detection under the Magnifying Glass
Molinari, A. C., Martini, T., Banov, L., Ierardi, A., Leotta, M., Strangio, A., & Santoro, R. C. (2023) · Journal of Clinical Medicine · doi:10.3390/jcm12206654 · Cited by 9 · see citing works
Shimonishi, N., Ogiwara, K., Oda, Y., Kawabe, T., Emmi, M., Shima, M., & Nogami, K. (2021) · Thrombosis and Haemostasis · doi:10.1055/s-0040-1721776 · Cited by 8 · see citing works
Analytical dilemmas in lupus anticoagulant detection
Moore, G. W. (2023) · Exploration of Immunology · doi:10.37349/ei.2023.00104 · Cited by 5 · see citing works
Self-citationShimomura, D., Kumano, O., Ueda, K., Kitano, K., Arai, N., Shimada, M., & Kamioka, M. (2024) · International Journal of Laboratory Hematology · doi:10.1111/ijlh.14289 · Cited by 5 · see citing works
“Although many studies had compared the sensitivity among the mixing test indexes in LA-positive samples, 10,14 there were a few studies to investigate the specificity in non-LA samples, especially for samples of LA and progressive coagulation factor inhibitor from acquired hemophilia A. 15 – 17…”
Self-citationLing, L., Liu, C., Huang, X., Liao, J., Jia, J., & Zhou, J. (2022) · International Journal of Laboratory Hematology · doi:10.1111/ijlh.13830 · Cited by 5 · see citing works
Lupus Anticoagulant Testing: Dilute Prothrombin Time (dPT)
Moore, G. W. (2023) · Methods in molecular biology · doi:10.1007/978-1-0716-3175-1_17 · Cited by 5 · see citing works
Self-citationMatsuda, M., Hoshiyama, Y., Ogawa, K., Emmi, M., Terai, S., & Moriyama, M. (2023) · Research and Practice in Thrombosis and Haemostasis · doi:10.1016/j.rpth.2023.100065 · Cited by 4 · see citing works
“APTT mixing tests, without incubation, were performed in duplicate in the prepared samples, using an automated coagulation analyzer (CP3000, SEKISUI MEDICAL) and an LA-sensitive reagent, Thrombocheck APTT-SLA (Sysmex) [6].”
BackgroundMethodInfluentialComparison of different algorithms for lupus anticoagulant detection: a single-center experience
Apipongrat, D., Lamool, R., Arnutti, P., Ruangpratheep, C., & Chantkran, W. (2024) · Research and Practice in Thrombosis and Haemostasis · doi:10.1016/j.rpth.2024.102333 · Cited by 4 · see citing works
Matsuda, M., Shimomura, D., Suzuki, T., Tabuchi, Y., Kurono, H., Nishi, K., Arai, N., Hoshiyama, Y., Kamioka, M., Moriyama, M., et al. (2025) · Scientific Reports · doi:10.1038/s41598-025-15089-3 · Cited by 4 · see citing works
Kumano, O., Peyrafitte, M., & Amiral, J. (2023) · Exploration of Immunology · doi:10.37349/ei.2023.00110 · Cited by 3 · see citing works
Self-citationMarco, A. (2025) · Journal of Clinical Medicine · doi:10.3390/jcm14082791 · Cited by 2 · see citing works
“This cutoff can also be provided by the supplier; it is normally verified in 20–40 healthy subjects, and it is assumed that an adequate statistical study has been established for its determination [6,18].”
Kumano, O., Amiral, J., Dunois, C., Peyrafitte, M., & Moore, G. W. (2021) · International Journal of Laboratory Hematology · doi:10.1111/ijlh.13571 · Cited by 2 · see citing works
Self-citationPolice, P., Noikongdee, P., Phojanasenee, T., & Apipongrat, D. (2026) · International Journal of Laboratory Hematology · doi:10.1111/ijlh.70064 · Cited by 1 · see citing works
Kumano, O., & Moore, G. W. (2023) · Thrombosis Research · doi:10.1016/j.thromres.2023.09.014
Self-citationKamali, S., Dave, M., Raheja, P., Sivapalaratnam, S., & Platton, S. (2025) · International Journal of Laboratory Hematology · doi:10.1111/ijlh.14537
“…[12] Ratio = (Patient 1:1 APTT) (Normal Pool APTT) Compare the patient 1:1 APTT result with the upper limit of the reference range [19] 6 Compare the patient 1:1 APTT result to a mixing test cut-off derived from 1:1 APTT results of normal donors [20, 21] Note: Adapted from Adcock DM et al. [12].”
Imataki, O., Takahata, K., Mukai, Y., et al. (2025) · Cureus · doi:10.7759/cureus.97307
“The circulation anticoagulant index (ICA) [12], the normalized ratio [13], and the Mix-NPP [14] are specifically produced indices that normalize the results of the anti-lupus coagulant mixing test.”
Gatt, A., Riva, N., Sammut, L., Vella, K., Kitchen, S., Hickey, K. P., & Borg-Aquilina, D. (2026) · Seminars in Thrombosis and Hemostasis · doi:10.1055/a-2971-6729
- Authors
- Kumano, O., & Moore, G. W.
- Affiliations
- Sysmex (Japan) (JP)
- St Thomas' Hospital (GB)
- Corresponding author
- Kumano, Osamu
- Venue
- Research and Practice in Thrombosis and Haemostasis
- Publisher
- Elsevier BV
- Published
- 2018-01
- Volume
- 2
- Issue
- 1
- Pages
- 105-113
- ISSN
- 2475-0379 (ISSN-L)
- PMC
- PMC6055558
- OpenAlex
- W2779707539
- Cited by
- 26
- References
- 44
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 2 · 2023: 6 · 2022: 4 · 2021: 2 · 2019: 5 · 2018: 1 · 2017: 1
- FWCI
- 1.47
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Research and Practice in Thrombosis and Haemostasis (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (submittedVersion, cc-by-nc-nd)
- PubMed Central (submittedVersion, cc-by-nc-nd)
- Topics
- Health Sciences › Medicine › Rheumatology › Systemic Lupus Erythematosus Research
- Health Sciences › Medicine › Hematology › Blood Coagulation and Thrombosis Mechanisms
- Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
- Keywords
- lupus anticoagulant, activated partial thromboplastin time, antiphospholipid antibodies
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