Interference of Heterophilic Antibody in D-dimer Determination in an Asymptomatic Elderly Woman

Valverde Sara

Laboratory Medicine, Madonna della Navicella Hospital Chioggia, Venice, Italy.

Masiero Elena

Laboratory Medicine, Madonna della Navicella Hospital Chioggia, Venice, Italy.

Seguso Mara

Laboratory Medicine, Dell’Angelo Hospital Mestre, Venice, Italy.

Giordano Martina

Laboratory Medicine, Dell’Angelo Hospital Mestre, Venice, Italy.

Inglese Margherita

Internal Medicine, Madonna della Navicella Hospital Chioggia, Venice, Italy.

Gessoni Gianluca *

Transfusion Medicine, Dell’Angelo Hospital Mestre, Venice, Italy.

*Author to whom correspondence should be addressed.


Background: D-Dimer is considered a pivotal biomarker in diagnosis of disseminated intravascular coagulation and in differential diagnosis of thrombosis and pulmonary embolism.

Case Summary: BL, Caucasian woman, 81 years old, was admitted to hospital, in October 2023, for concussive head trauma after an accidental fall. The patient had a "non-assayable D-Dimer due to excess antigen" utilizing Sysmex Innovance D-dimer using a Sysmex CS 5100 analyser. This abnormal result was firstly observed in March 2022. A second Laboratory confirmed the raised D-dimer concentration. The patient had undergone periodic D-dimer checks which had always confirmed the results and had been treated with a direct FXa inhibitor.

Methods:  Patient’s samples were tested for D-dimer using different assays and different analysers, moreover sample diluted in phosphate buffer and heterophilic antibodies blocking reagent have been tested.

Results: The Sysmex Innovance D-dimer assay gave us, constantly “non-assayable D-dimer due to excess antigen" results; the HemosIL D-dimer HS assay gave us, constantly a raised D-dimer concentration (four to five higher than upper reference values); the Quidel Triage D-dimer gave us, constantly  D-dimer normal concentration. Results obtained from dilution curves confirmed the presence of high concentration high avidity  heterophilic antibodies.

Conclusions: Reports regarding the influence of heterophilic antibodies on the measurement of D‐dimer are quite uncommon in literature however, they constitute a significant potential risk. Interference from heterophile antibodies often has a different impact using different instruments and methods in the measurement of D‐dimer. Using a combination of different assays and analysers, of dilution strategy with heterophilic antibody blockers, and combining laboratory results with clinical examinations and imaging data, we were able to identify the interference and exclude the presence of thrombosis.

Keywords: D‐dimer, D‐dimer assays, heterophilic antibody, immunoassay, interference

How to Cite

Sara, V., Elena , M., Mara , S., Martina, G., Margherita, I., & Gianluca , G. (2024). Interference of Heterophilic Antibody in D-dimer Determination in an Asymptomatic Elderly Woman. International Blood Research & Reviews, 15(2), 1–9.


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Weitz JI, Fredenburgh JC, Eikelboom JW. A test in contest: D-dimer. J Am Coll Cardiol. 2017;70:2411–20. DOI: 10.1016/j.jacc.2017.09.024

Brotman DJ. Higher D-dimer thresholds for excluding pulmonary embolism: No free lunch? Ann Intern Med. 2022;175:295-296. DOI: 10.7326/M21-4295

Udovenko A, Makogonenko Y, Korolova D, et al. Formation and elimination of soluble fibrin and D-dimer in the bloodstream. Croat Med J. 2023;64:421-429. DOI: 10.3325/cmj.2023.64.421

Riley RS, Gilbert AR, Dalton JB, et al. Widely used types and clinical applications of D-dimer assay. Lab Med. 2016;47: 90–102. DOI: 10.1093/labmed/lmw001

Talon L, Fourneyron V, Trapani A, et al. Analytical performance of a new immune turbidimetric D-dimer assay and comparison with available assays. Res Pract Thromb Haemost. 2022;6:e12660. DOI: 10.1002/rth2.12660.

Favresse J, Lippi G, Roy PM, et al. D-dimer: Preanalytical, analytical, postanalytical variables, and clinical applications. Crit Rev Clin Lab Sci. 2018; 55:548-577. DOI: 10.1080/10408363.2018.1529734

Mastella AK, de Carvalho JAM, Stein CS, et al. Interference of icterus on plasma D-dimer levels measured using immunoturbidimetric assays. Blood Coagul Fibrinolysis. 2021;32:162-163. DOI: 10.1097/MBC.0000000000001004

Tate J, Ward G. Interferences in immunoassay. Clin Biochem Rev. 2004; 25:105–20.

Cheng X, Guo X, Chai X, et al. Heterophilic antibody interference with TSH measurement on different immunoassay platforms. Clin Chim Acta. 2021;512:63-65. DOI: 10.1016/j.cca.2020.11.018

Zhang XY, Zhang XX, Xu JL, et al. Identification of and solution for false D-dimer results. J Clin Lab Anal. 2020;34: e23216. DOI: 10.1002/jcla.23216

Wells PS, Anderson DR, Rodger M, et al. Derivation of a simple clinical model to categorize patients probability of pulmonary embolism: increasing the models utility with the Simpli RED D-dimer. Thromb Haemost. 2000;83:416–20. DOI: 10.1055/s-0037-1613830

Wells PS, Anderson DR, Bormanis J, et al. Value of assessment of pretest probability of deep-vein thrombosis in clinical management. Lancet. 1997;350(9094): 1795–8.

DOI: 10.1016/S0140-6736(97)08140-3

Adriaansen MJ, Morison IM, Perry HE. Discrepancies between two D-dimer assays and impact on clinical decisions; a retrospective analysis of samples tested in community and hospital-based laboratories in Auckland. N Z Med J. 2024;137: 12-19. DOI: 10.26635/6965.6302

Ma H, Chen CH, Li YM, Wang JJ, Hu ZD. Falsely elevated D-dimer partially caused by heterophilic antibodies: A case report. Clin Chim Acta. 2022; 535(1):40-142. DOI: 10.1016/j.cca.2022.08.011

Zhang L, Bao G, Zheng Z. Two cases of pseudoelevation of plasma D-dimer levels. Clin Lab. 2021;67(9). DOI: 10.7754/Clin.Lab.2021.210203

Chen L, Chen Y. Performance Evaluation of the Sysmex CS-5100 Automated Coagulation Analyzer. Clin Lab. 2015; 61:653-60. DOI: 10.7754/clin.lab.2014.141124

Favaloro EJ, Mohammed S, Vong R, et al. Verification of the ACL Top 50 Family (350, 550, and 750) for harmonization of routine coagulation assays in a large network of 60 laboratories. Am J Clin Pathol. 2021; 156:661-678. DOI: 10.1093/ajcp/aqab004.

Ghys T, Achtergael W, Verschraegen I,et al. Diagnostic accuracy of the Triage D-dimer test for exclusion of venous thromboembolism in outpatients. Thromb Res. 2008;121:735-41. DOI: 10.1016/j.thromres.2007.07.012

Anderson LD Jr, Bladé J, Kyle RA. A case of benign immunoglobulin D monoclonal gammopathy of undetermined significance with 26 years of follow-up. E J Haem. 2024;5:235-237.

DOI: 10.1002/jha2.846

Sun L, Wang N, Feng Y, et al. The distribution of heterophilic antigens and their relationship with autoimmune diseases. Front Immunol. 2023;14: 12756-58. DOI :10.3389/fimmu.2023.1275658

Aliberti L, Gagliardi I, Dorizzi RM, et al. Hypeprolactinemia: Still an insidious diagnosis. Endocrine. 2021;72:928-931 DOi: 10.1007/s12020-020-02497-w

Hammarsten O, Becker C, Engberg AE. Methods for analyzing positive cardiac troponin assa interference. Clin Biochem. 2023;116:24-30. DOI: 10.1016/j.clinbiochem.2023.03.004

Zhao W, Duan L, Fang L, et al. Persistent increase of carbohydrate antigen 19-9 with an unknown reason: A seven-year follow-up case. J Clin Lab Anal. 2022;36:e24792. DOI: 10.1002/jcla.24792

Carter-Brzezinski L, Houston S, Thachil J. D-dimers: A most misunderstood test. Br J Hosp Med (Lond). 2021;82:1-5. DOI: 10.12968/hmed.2021.0279

Lippi G, Avanzini P, Zobbi V, Ippolito L. Influence of mechanical hemolysis of blood on two D-dimer immunoassays. Blood Coagul Fibrinolysis. 2012;23:461-3. DOI: 10.1097/MBC.0b013e3283549696

Negrini D, Bernardi D, Antonelli G, Plebani M. Interference of lipemia in samples for routine coagulation testing using a Sysmex CS-5100 coagulometer. Int J Lab Hematol. 2019;41:772-777.

DOI: 10.1111/ijlh.13108

Robier C, Edler E, Klescher D, Neubauer M. False-positive D-dimer result in a latex-enhanced immunoassay caused by interfering human anti-mouse antibodies. Clin Chem Lab Med. 2014;52:253-5. DOI: 10.1515/cclm-2014-0496

Wu Y, Xiao YX, Huang TY, et al. What makes D-dimer assays suspicious-heterophilic antibodies? J Clin Lab Anal. 2019;33:e22687. DOI: 10.1002/jcla.22687.

Ozbalci D, Doguc DK, Yilmaz G, et al. Interference of D-dimer levels from heterophilic antibody in Covid-19: A serious concern in treatment and follow-up of patients. Int J Lab Hematol. 2022;44:e13-e16. DOI: 10.1111/ijlh.13654

Lippi G, Ippolito L, Tondelli MT, Favaloro EJ. Interference from heterophilic antibodies in D-dimer assessment. A case report. Blood Coagul Fibrinolysis. 2014;25:277-9. DOI: 10.1097/MBC.0000000000000017

Favaloro EJ, Dean E. Variability in D-dimer reporting revisited. Pathology. 2021;53:538-540.

DOI: 10.1016/j.pathol.2020.08.010.

Sun HX, Ge H, Xu ZQ, Sheng HM. Clinical laboratory investigation of a patient with an extremely high D-dimer level: A case report. World J Clin Cases. 2020;8:3560-3566. DOI: 10.12998/wjcc.v8.i16.3560

Lippi G, Aloe R, Meschi T, ei al. Interference from heterophilic antibodies in troponin testing. Case report and systematic review of the literature. Clin Chim Acta. 2013;426:79-84. DOI: 10.1016/j.cca.2013.09.004