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
Abstract
BACKGROUND: Phospholipid-dependent coagulation tests for lupus anticoagulant (LA) are considered an important step for the diagnosis of anti-phospholipid syndrome; however, LA laboratory detection is difficult because of many variables. Five hospital laboratories, located in a North-Italy area and using the same method for LA testing, cooperated to standardise sample treatment and analytical procedure in order to define the upper values for LA negativity. METHODS: In total, 200 normal subjects (40 for each centre) were studied for six LA functional assays, using the same procedure, reagent lot and analyser type. The first tests done were LA screen and LA confirm assays, based on diluted Russell's Viper Venom Time, with low and high phospholipid content, respectively. The second tests performed were silica clotting time screen and confirm assays, based on activated partial thromboplastin time, with low and high phospholipid content, respectively. Finally, two mixing assays were executed for both screening assays, diluting patient sample with a pool prepared with plasma collected from the study population. RESULTS: Data analysis demonstrated a difference between centres for all assays when results were expressed in seconds; the difference disappeared when results are normalised with the local mean normal value of each centre and are expressed as a normalised ratio. The study population was normally distributed; so the value corresponding to 99th percentile was used as limit value for LA negativity. Values expressed as normalised ratio, for LA and silica clotting time screenings were 1.22 and 1.23, respectively. CONCLUSIONS: The study allowed us to define a uniform approach to LA testing and evaluation for laboratories employing the same methods.
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Reagent lot variation is a probable explanation for the differences, supported by a multicenter study in which every center used one common lot.
“Probably, they might be explained by the variation in reagent lots, as confirmed in another multicenter study with the same reagent lot used by all participating centers.77”
Some works have shown that the values from the parametric approach do not respect the Gaussian probability model.
“(even if some works have shown that these values do not respect the Gaussian probability model [61,62])”
From Lupus Anticoagulant Detection under the Magnifying Glass (2023) · cites this paper as [62]
Cites
MIYAKIS, S., LOCKSHIN, M. D., ATSUMI, T., BRANCH, D. W., BREY, R. L., CERVERA, R., DERKSEN, R. H. W. M., DE GROOT, P. G., KOIKE, T., MERONI, P. L., et al. (2006) · Journal of Thrombosis and Haemostasis · doi:10.1111/j.1538-7836.2006.01753.x · Cited by 7,203 · see citing works
Update of the guidelines for lupus anticoagulant detection
PENGO, V., TRIPODI, A., REBER, G., RAND, J. H., ORTEL, T. L., GALLI, M., & DE GROOT, P. G. (2009) · Journal of Thrombosis and Haemostasis · doi:10.1111/j.1538-7836.2009.03555.x · Cited by 1,197 · see citing works
Brandt, J. T., Triplett, D. A., Alving, B. M., & Scharrer, I. (1995) · PubMed · Cited by 1,164 · see citing works
Brandt, J. T., Triplett, D. A., Alving, B., & Scharrer, I. (1995) · Thrombosis and Haemostasis · doi:10.1055/s-0038-1649901 · Cited by 993 · see citing works
Galli, M., Luciani, D., Bertolini, G., & Barbui, T. (2003) · Blood · doi:10.1182/blood-2002-02-0441 · Cited by 974 · cited 1× · see citing works
“Early in the last decade, the presence of LA, detected through coagulation assays, has shown to be a stronger thrombotic risk factor than other antiphospholipid antibodies [12] , thus leading LA determination to a pivotal role in the laboratory diagnosis of APS [13, 14] .”
BackgroundRuiz‐Irastorza, G., Crowther, M. A., Branch, W. D., & Khamashta, M. A. (2010) · The Lancet · doi:10.1016/s0140-6736(10)60709-x · Cited by 864 · see citing works
Guidelines on the investigation and management of antiphospholipid syndrome
Keeling, D., Mackie, I., Moore, G. W., Greer, I. A., Greaves, M., & British Committee for Standards in Haematology (2012) · British Journal of Haematology · doi:10.1111/j.1365-2141.2012.09037.x · Cited by 740 · see citing works
The antiphospholipid syndrome: ten years on
Hughes, G. R. V. (1993) · The Lancet · doi:10.1016/0140-6736(93)91477-4 · Cited by 628 · see citing works
Guidelines On The Investigation and Management Of The Antiphospholipid Syndrome
Greaves, M., Cohen, H., Machin, S. J., & Mackie, I. J. (2000) · British Journal of Haematology · doi:10.1046/j.1365-2141.2000.02069.x · Cited by 294 · see citing works
Exner, T., Triplett, D. A., Taberner, D. A., & Machin, S. J. (1991) · PubMed · Cited by 243 · see citing works
Guidelines for Testing and Revised Criteria for Lupus Anticoagulants
Exner, T., Triplett, D. A., Taberner, D. A., & Machin, S. J. (1991) · Thrombosis and Haemostasis · doi:10.1055/s-0038-1648143 · Cited by 203 · see citing works
Detection and Quantitative Evaluation of Lupus Circulating Anticoagulant Activity
Rosner, E., Pauzner, R., Lusky, A., Modan, M., & Many, A. (1987) · Thrombosis and Haemostasis · doi:10.1055/s-0038-1651083 · Cited by 188 · see citing works
Pierangeli, S., de Groot, P. G., Dlott, J. S., Favaloro, E. J., Harris, E., Lakos, G., Ortel, T. L., Meroni, P. L., Otomo, K., Pengo, V., et al. (2011) · Lupus · doi:10.1177/0961203310395055 · Cited by 179 · see citing works
Laboratory Investigation of Thrombophilia
Tripodi, A., & Mannucci, P. M. (2001) · Clinical Chemistry · doi:10.1093/clinchem/47.9.1597 · Cited by 130 · see citing works
Antiphospholipid Syndrome: Diagnostic Aspects of Lupus Anticoagulants
Arnout, J. (2001) · Thrombosis and Haemostasis · doi:10.1055/s-0037-1616204 · Cited by 121 · see citing works
A protocol for determination of anticardiolipin antibodies by ELISA
Pierangeli, S. S., & Harris, E. N. (2008) · Nature Protocols · doi:10.1038/nprot.2008.48 · Cited by 106 · see citing works
Survey of lupus anticoagulant diagnosis by central evaluation of positive plasma samples
Pengo, V., Biasiolo, A., Gresele, P., Marongiu, F., Erba, N., Veschi, F., Ghirarduzzi, A., Candia, E. D., Montaruli, B., Testa, S., et al. (2007) · Journal of Thrombosis and Haemostasis · doi:10.1111/j.1538-7836.2007.02454.x · Cited by 95 · see citing works
Laboratory Testing for Lupus Anticoagulants: A Review of Issues Affecting Results
Tripodi, A. (2007) · Clinical Chemistry · doi:10.1373/clinchem.2007.089524 · Cited by 81 · see citing works
Biasiolo, A., Rampazzo, P., Brocco, T., & Pengo, V. (1999) · Thrombosis and Haemostasis · doi:10.1055/s-0037-1614453 · Cited by 76 · see citing works
Jennings, I., Greaves, M., Mackie, I. J., Kitchen, S., Woods, T. A. L., Preston, F. E., & ation, O. B. O. T. U. N. E. Q. A. S. F. B. C. (2002) · British Journal of Haematology · doi:10.1046/j.1365-2141.2002.03821.x · Cited by 72 · see citing works
A “Percent Correction” Formula for Evaluation of Mixing Studies
Chang, S.-H., Tillema, V., & Scherr, D. (2002) · American Journal of Clinical Pathology · doi:10.1309/rrek-8l6m-d2kc-hwlh · Cited by 67 · cited 1× · see citing works
“In these guidelines a three-step strategy is recommended: a) screening test (with a low phospholipid concentration) to verify a prolongation of a phospholipiddependent clotting time beyond the upper limit of the reference range; b) mixing test to exclude a coagulation factor deficiency [22, 23] ; c) confirmation test…”
MethodConsensus guidelines on anti-cardiolipin antibody testing and reporting
Wong, R. C. W., Gillis, D., Adelstein, S., Baumgart, K. W., Favaloro, E. J., Hendle, M. J., Homes, P., Pollock, W., Smith, S., Steele, R. H., et al. (2004) · Pathology · doi:10.1080/00313020310001643615 · Cited by 63 · see citing works
Pengo, V., Banzato, A., Bison, E., Bracco, A., Denas, G., & Ruffatti, A. (2012) · Seminars in Thrombosis and Hemostasis · doi:10.1055/s-0032-1304719 · Cited by 59 · see citing works
Chantarangkul, V., Tripodi, A., Arbini, A., & Mannucci, P. M. (1992) · Thrombosis Research · doi:10.1016/0049-3848(92)90266-d · Cited by 52 · see citing works
Interpretation of normal plasma mixing studies in the laboratory diagnosis of lupus anticoagulants
Devreese, K. M. J. (2007) · Thrombosis Research · doi:10.1016/j.thromres.2006.03.012 · Cited by 46 · see citing works
European attempts for the standardisation of the antiphospholipid antibodies
Tincani, A., Filippini, M., Scarsi, M., Galli, M., & Meroni, P. L. (2009) · Lupus · doi:10.1177/0961203309106919 · Cited by 40 · see citing works
Consensus guidelines on anti-beta 2 glycoprotein I testing and reporting
Wong, R. C. W., Favaloro, E. J., Adelstein, S., Baumgart, K. W., Bird, R. J., Brighton, T., Empson, M. B., Gillis, D., Hendle, M. J., Laurent, R., et al. (2007) · Pathology · doi:10.1080/00313020701717720 · Cited by 37 · see citing works
Favaloro, E. J., & Wong, R. C. W. (2011) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm.2011.064 · Cited by 36 · see citing works
Galli, M. (2008) · Seminars in Thrombosis and Hemostasis · doi:10.1055/s-0028-1085474 · Cited by 32 · see citing works
Žigon, P., Ambrožič, A., Čučnik, S., Kveder, T., Rozman, B., & Božič, B. (2011) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm.2011.162 · Cited by 25 · see citing works
Antiphospholipid syndrome: interpretation of laboratory data
Pengo, V. (2010) · Journal of Thrombosis and Haemostasis · doi:10.1111/j.1538-7836.2010.04151.x · Cited by 15 · see citing works
Grypiotis, P., Ruffatti, A., Pengo, V., et al. (2006) · Journal of Clinical Laboratory Analysis · doi:10.1002/jcla.20093 · Cited by 14 · see citing works
The epidemiology of the antiphospholipid syndrome: Who is at risk?
Finazzi, G. (2001) · Current Rheumatology Reports · doi:10.1007/s11926-001-0030-5 · Cited by 14 · see citing works
Froom, P., & Barak, M. (2011) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm.2011.768 · Cited by 9 · see citing works
Mameli, A., Barcellona, D., Vannini, M. L., & Marongiu, F. (2011) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm.2011.111 · Cited by 7 · see citing works
Montaruli, B., Vaccarino, A., Foli, C., Rus, C., Agnes, C., Saitta, M., & Bazzan, M. (2005) · Clinical Chemistry · doi:10.1373/clinchem.2004.042028 · Cited by 2 · see citing works
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Devreese, K. M. J., de Groot, P. G., de Laat, B., Erkan, D., Favaloro, E. J., Mackie, I., Martinuzzo, M., Ortel, T. L., Pengo, V., Rand, J. H., et al. (2020) · Journal of Thrombosis and Haemostasis · doi:10.1111/jth.15047 · Cited by 439 · see citing works
Antiphospholipid antibody testing and standardization
Devreese, K. M. J. (2014) · International Journal of Laboratory Hematology · doi:10.1111/ijlh.12234 · Cited by 98 · see citing works
Moore, G. W., Jones, P. O., Platton, S., Hussain, N., White, D., Thomas, W., Rigano, J., Pouplard, C., Gray, E., & Devreese, K. M. J. (2021) · Journal of Thrombosis and Haemostasis · doi:10.1111/jth.15438 · Cited by 47 · see citing works
Moore, G. W., Culhane, A. P., Daw, C. R., Noronha, C. P., & Kumano, O. (2016) · Thrombosis Research · doi:10.1016/j.thromres.2016.01.019 · Cited by 42 · see citing works
Current Controversies in Lupus Anticoagulant Detection
Moore, G. (2016) · Antibodies · doi:10.3390/antib5040022 · Cited by 36 · see citing works
Tripodi, A., Chantarangkul, V., Cini, M., Devreese, K., Dlott, J. S., Giacomello, R., Gray, E., Legnani, C., Martinuzzo, M. E., Pradella, P., et al. (2017) · Journal of Thrombosis and Haemostasis · doi:10.1111/jth.13678 · Cited by 35 · see citing works
Self-citationMoore, G. W., & Kumano, O. (2020) · Journal of Thrombosis and Haemostasis · doi:10.1111/jth.14669 · Cited by 18 · see citing works
Lupus 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
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
- Authors
- Pradella, P., Azzarini, G., Santarossa, L., Caberlotto, L., Bardin, C., Poz, A., D’Aurizio, F., & Giacomello, R.
- Affiliations
- Ospedale di Cattinara (IT)
- Ospedale dell' Angelo (IT)
- Azienda Ospedaliera Santa Maria Degli Angeli (IT)
- Ca' Foncello Hospital (IT)
- University of Udine (IT)
- Corresponding author
- Pradella, Paola
- Venue
- Clinical Chemistry and Laboratory Medicine (CCLM)
- Publisher
- Walter de Gruyter GmbH
- Published
- 2012-09-22
- Volume
- 51
- Issue
- 2
- Pages
- 379-385
- ISSN
- 1434-6621 (ISSN-L)
- 1437-4331
- OpenAlex
- W1994162452
- Cited by
- 26
- References
- 36
- Citations by year
- 2026: 1 · 2023: 2 · 2022: 3 · 2021: 2 · 2020: 2 · 2019: 1 · 2018: 1 · 2017: 2 · 2016: 2 · 2015: 3 · 2014: 3 · 2013: 3 · 2012: 1
- FWCI
- 2.45
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Clinical Chemistry and Laboratory Medicine (CCLM) (publishedVersion)
- PubMed (publishedVersion)
- Institutional Research Information System (University of Udine) (submittedVersion)
- 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, antiphospholipid syndrome, reference limits, healthy population, multicentre collaboration, Russell's viper venom time, activated partial thromboplastin time
- MeSH terms
- Adolescent, Adult, Aged, Blood Coagulation Tests, Female, Humans, Male, Middle Aged, Reference Values, Multicenter Studies as Topic, Lupus Coagulation Inhibitor, Antiphospholipid Syndrome, Young Adult
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