FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Journal of veterinary internal medicine2023; 38(1); 187-196; doi: 10.1111/jvim.16888

Immunoreactivity of canine, feline, and equine D-dimer with antibodies to human D-dimer.

Abstract: Commercially available D-dimer assays use antibodies against human D-dimer, with limited sensitivity and specificity data in companion animals. Objective: To evaluate the immunoreactivity of D-dimer in plasma of dogs, horses, and cats with commercially available antibodies to human D-dimer. Methods: Plasma samples were collected from healthy dogs and horses, and from surplus feline plasma submitted for diagnostic purposes. Methods: Descriptive research study. A cross-linked fibrin lysate was prepared from plasma samples, and SDS-PAGE and immunoblotting were performed with a variety of commercially available antibodies to human D-dimer. Results: The selected antibodies demonstrated variable reactivity with D-dimer of each species. The monoclonal antibody DD44 bound canine D-dimer with good specificity and sensitivity, but this antibody did not react with feline or equine D-dimer. The polyclonal antibody D2D bound putative D-dimer in dogs, cats, and horses with good specificity, and higher sensitivity compared to human D-dimer. Conclusions: The variable performance of commercially available human D-dimer assays between species is, in part, because of inter-species variation in D-dimer immunoreactivity. The use of these assays should follow validation studies. Monoclonal antibody DD44 could be a focus for the development of a canine-specific assay.
© 2023 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.
Get Full Text
Publication Date: 2023-11-10 PubMed ID: 37950415PubMed Central: PMC10800179DOI: 10.1111/jvim.16888Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

Overview

  • Researchers investigated how well antibodies designed to detect human D-dimer react with D-dimer proteins found in dogs, cats, and horses.
  • The goal was to assess the suitability of existing human D-dimer tests for measuring these proteins in common companion animals.

Background

  • D-dimer is a protein fragment produced when blood clots are broken down in the body.
  • Assays measuring D-dimer are commonly used in human medicine to diagnose conditions related to abnormal blood clotting.
  • Commercial D-dimer test kits are usually optimized for human D-dimer and use antibodies specific to human D-dimer proteins.
  • There is limited information on the effectiveness of these human-based tests when applied to veterinary species such as dogs, cats, and horses.

Objectives

  • To evaluate the immunoreactivity (binding affinity) of canine, feline, and equine D-dimer proteins with several commercially available antibodies developed against human D-dimer.
  • To determine whether these antibodies show sufficient sensitivity and specificity to detect D-dimer in these animal species.
  • To identify any antibodies potentially useful for developing species-specific D-dimer tests in animals.

Methods

  • Plasma samples were obtained from:
    • Healthy dogs and horses.
    • Surplus feline plasma collected for diagnostic purposes.
  • A cross-linked fibrin lysate (enriched in D-dimer) was prepared from the plasma samples.
  • SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and immunoblotting techniques were used to detect D-dimer proteins.
  • Multiple commercially available antibodies against human D-dimer were tested for their ability to bind D-dimer from each species.

Key Findings

  • Antibodies showed variable reactivity depending on the animal species.
    • Monoclonal antibody DD44:
      • Bound canine D-dimer effectively with good specificity and sensitivity.
      • Did not react with feline or equine D-dimer.
    • Polyclonal antibody D2D:
      • Bound D-dimer in dogs, cats, and horses with good specificity.
      • Displayed even higher sensitivity in these species compared to reactivity with human D-dimer.

Conclusions and Implications

  • There are important differences in how D-dimer proteins from different species react with antibodies against human D-dimer.
  • This variability contributes to inconsistent performance of human D-dimer assays when applied to veterinary species.
  • Veterinary use of human D-dimer tests should be accompanied by appropriate validation in each species to ensure accurate results.
  • The monoclonal antibody DD44 shows promise for developing a canine-specific D-dimer assay due to its focused and sensitive binding to canine D-dimer.
  • Polyclonal antibodies like D2D might serve as broad detection tools across multiple species but may lack the specificity of monoclonal antibodies.

Significance

  • This study informs veterinary diagnostic laboratories and clinicians about the limitations and potential of human D-dimer assays in animals.
  • It supports efforts to develop or validate species-specific D-dimer tests, improving clinical diagnosis of clotting disorders in companion animals.

Cite This Article

APA
Brown JE, Noormohammadi AH, Courtman NF. (2023). Immunoreactivity of canine, feline, and equine D-dimer with antibodies to human D-dimer. J Vet Intern Med, 38(1), 187-196. https://doi.org/10.1111/jvim.16888

Publication

Journal of veterinary internal medicine
ISSN: 1939-1676
NlmUniqueID: 8708660
Country: England
Language: English
Volume: 38
Issue: 1
Pages: 187-196

Researcher Affiliations

Brown, Juliet E
  • Faculty of Veterinary and Agricultural Sciences, University of Melbourne, Melbourne, Victoria, Australia.
Noormohammadi, Amir H
  • Faculty of Veterinary and Agricultural Sciences, University of Melbourne, Melbourne, Victoria, Australia.
Courtman, Natalie F
  • Faculty of Veterinary and Agricultural Sciences, University of Melbourne, Melbourne, Victoria, Australia.

MeSH Terms

  • Humans
  • Cats
  • Animals
  • Dogs
  • Horses
  • Fibrin Fibrinogen Degradation Products
  • Antibodies, Monoclonal
  • Plasma
  • Sensitivity and Specificity

Grant Funding

  • 2021 / American Society for Veterinary Clinical Pathology
  • W21-048 / EveryCat Health Foundation
  • University of Melbourne

Conflict of Interest Statement

Authors declare no conflict of interest.

References

This article includes 42 references
  1. Weisel JW, Dempfle CEH. Fibrinogen structure and function.. Hemostasis and Thrombosis: Basic Principles and Clinical Practice 6th ed. Philadelphia: Wolters Kluwer; 2012:867.
  2. Song KS, Kim YA, Kim HK, Park Q. Incidence and possible reasons for discordant results between positive FDP and negative D‐dimer latex assays in clinical specimens.. Yonsei Med J 1999;40(2):107‐111.
    doi: 10.3349/ymj.1999.40.2.107pubmed: 10333712google scholar: lookup
  3. Carretón E, Morchón R, González‐Miguel J, Simón F, Juste MC, Montoya‐Alonso JA. Variation of d‐dimer values as assessment of pulmonary thromboembolism during adulticide treatment of heartworm disease in dogs.. Vet Parasitol 2013;195(1–2):106‐111.
    doi: 10.1016/j.vetpar.2013.01.005pubmed: 23384581google scholar: lookup
  4. Nelson OL, Andreasen C. The utility of plasma D‐dimer to identify thromboembolic disease in dogs.. J Vet Intern Med 2003;17(6):830‐834.
    doi: 10.1892/0891-6640(2003)017<0830:tuopdt>2.3.co;2pubmed: 14658720google scholar: lookup
  5. Laforcade AM, Freeman LM, Shaw SP, Brooks MB, Rozanski EA, Rush JE. Hemostatic changes in dogs with naturally occurring sepsis.. J Vet Intern Med 2003;17(5):674‐679.
    doi: 10.1111/j.1939-1676.2003.tb02499.xpubmed: 14529134google scholar: lookup
  6. Kang MH, Heo RY, Park HM. Evaluation of D‐dimer concentrations in clinically ill dogs with high risk of thromboembolic disease.. Pak Vet J 2016;36(2):219‐223.
  7. Scott‐Moncrieff J, Treadwell N, McCullough S, Brooks M. Hemostatic abnormalities in dogs with primary immune‐mediated hemolytic anemia.. J Am Anim Hosp Assoc 2001;37(3):220‐227.
    doi: 10.5326/15473317-37-3-220pubmed: 11361112google scholar: lookup
  8. Ribera T, Monreal L, Armengou L, Ríos J, Prades M. Synovial fluid D‐dimer concentration in foals with septic joint disease.. J Vet Intern Med 2011;25(5):1113‐1117.
    doi: 10.1111/j.1939-1676.2011.0758.xpubmed: 21781162google scholar: lookup
  9. Cesarini C, Monreal L, Armengou L, Delgado MÁ, Ríos J, Jose‐Cunilleras E. Association of Admission Plasma D‐dimer concentration with diagnosis and outcome in horses with colic.. J Vet Intern Med 2010;24(6):1490‐1497.
    doi: 10.1111/j.1939-1676.2010.0618.xpubmed: 20946373google scholar: lookup
  10. Argüelles D, Delgado MA, Armengou L, Carmona JU, Prades M. Relationship between plasma and peritoneal fluid concentrations of D‐dimer and transforming growth factor Beta 1 in horses with colic.. J Equine Vet 2015;35(8):629‐635.
    doi: 10.1016/j.jevs.2015.06.002google scholar: lookup
  11. Delgado MA, Monreal L, Armengou L, Ríos J, Segura D. Peritoneal D‐dimer concentration for assessing peritoneal fibrinolytic activity in horses with colic.. J Vet Intern Med 2009;23(4):882‐889.
    doi: 10.1111/j.1939-1676.2009.0344.xpubmed: 19566853google scholar: lookup
  12. Monreal L, Angles AM, Monreal M, Espada Y, Monastierio J. Changes in haemostasis in endurance horses: detection by highly sensitive ELISA‐tests.. Equine Vet J 2010;27(S18):120‐123.
    doi: 10.1111/j.2042-3306.1995.tb04903.xgoogle scholar: lookup
  13. Antović J. ‘Point‐of‐Care’ D‐Dimer testing.. J Med Biochem 2010;29(4):282‐287.
    doi: 10.2478/v10011-010-0032-9google scholar: lookup
  14. Suzuki T, Distante A, Zizza A. Diagnosis of acute aortic dissection by D‐dimer.. Circulation 2009;119(20):2702‐2707.
    doi: 10.1161/circulationaha.108.833004pubmed: 19433758google scholar: lookup
  15. Epstein SE, Hopper K, Mellema MS, Johnson LR. Diagnostic utility of D‐dimer concentrations in dogs with pulmonary embolism.. J Vet Intern Med 2013;27(6):1646‐1649.
    doi: 10.1111/jvim.12177pubmed: 24033697google scholar: lookup
  16. Stokol T. Plasma D‐dimer for the diagnosis of thromboembolic disorders in dogs.. Vet Clin North Am Small Anim Pract 2003;33(6):1419‐1435.
    doi: 10.1016/s0195-5616(03)00096-2pubmed: 14664206google scholar: lookup
  17. Stokol T, Brooks MB, Erb HN, Mauldin GE. D‐dimer concentrations in healthy dogs.. Am J Vet Res 2000;61(4):393‐398.
    doi: 10.2460/ajvr.2000.61.393pubmed: 10772103google scholar: lookup
  18. Wada H, Sakuragawa N, Mori Y. Hemostatic molecular markers before the onset of disseminated intravascular coagulation.. Am J Hematol 1999;60(4):273‐278.
    doi: 10.1002/(sici)1096-8652(199904)60:4<273::aid-ajh4>3.0.co;2-npubmed: 10203100google scholar: lookup
  19. Griffin A, Callan MB, Shofer FS, Giger U. Evaluation of a canine D‐dimer point‐of‐care test kit for use in samples obtained from dogs with disseminated intravascular coagulation, thromboembolic disease, and hemorrhage.. Am J Vet Res 2003;64(12):1562‐1569.
    doi: 10.2460/ajvr.2003.64.1562pubmed: 14672437google scholar: lookup
  20. Dolente BA, Wilkins PA, Boston RC. Clinicopathologic evidence of disseminated intravascular coagulation in horses with acute colitis.. J Am Vet Med Assoc 2002;220(7):1034‐1038.
    doi: 10.2460/javma.2002.220.1034pubmed: 12420783google scholar: lookup
  21. Monreal L, Anglés A, Espada Y, Monasterio J, Monreal M. Hypercoagulation and hypofibrinolysis in horses with colic and DIC.. Equine Vet J 2000;32(S32):19‐25.
    doi: 10.1111/j.2042-3306.2000.tb05329.xpubmed: 11202377google scholar: lookup
  22. Welch RD, Watkins JP, Taylor TS, Cohen ND, Carter GK. Disseminated intravascular coagulation associated with colic in 23 horses (1984–1989).. J Vet Intern Med 1992;6(1):29‐35.
    doi: 10.1111/j.1939-1676.1992.tb00982.xpubmed: 1548623google scholar: lookup
  23. Tholen I, Weingart C, Kohn B. Concentration of D‐dimers in healthy cats and sick cats with and without disseminated intravascular coagulation (DIC).. J Feline Med Surg 2009;11(10):842‐846.
    doi: 10.1016/j.jfms.2009.04.008pmc: PMC11135504pubmed: 19539510google scholar: lookup
  24. Brazzell JL, Borjesson DL. Evaluation of plasma antithrombin activity and D‐dimer concentration in populations of healthy cats, clinically ill cats, and cats with cardiomyopathy.. Vet Clin Path 2007;36(1):79‐84.
    doi: 10.1111/j.1939-165x.2007.tb00186.xpubmed: 17311199google scholar: lookup
  25. Dempfle CE. Validation, calibration, and specificity of quantitative D‐dimer assays.. Seminars Vasc Med 2005;5(4):315‐320.
    doi: 10.1055/s-2005-922476pubmed: 16302152google scholar: lookup
  26. Török-Nagy B, Antal J, Dénes B. Generation and characterization of D‐dimer specific monoclonal antibodies for use in latex agglutination test.. PloS One 2019;14(2):e0212104.
    doi: 10.1371/journal.pone.0212104pmc: PMC6375587pubmed: 30763351google scholar: lookup
  27. Flatland B, Friedrichs KR, Klenner S. Differentiating between analytical and diagnostic performance evaluation with a focus on the method comparison study and identification of bias.. Vet Clin Path 2014;43(4):475‐486.
    doi: 10.1111/vcp.12199pubmed: 25257670google scholar: lookup
  28. Jennings L, Deerlin VMV, Gulley ML. Recommended principles and practices for validating clinical molecular pathology tests.. Arch Pathol Lab Med 2009;133(5):743‐755.
    doi: 10.5858/133.5.743pubmed: 19415949google scholar: lookup
  29. Cleophas T, Droogendijk J, van Ouwerkerk B. Validating diagnostic tests, correct and incorrect methods, new developments.. Curr Clin Pharmacol 2008;3(2):70‐76.
    doi: 10.2174/157488408784293697pubmed: 18690881google scholar: lookup
  30. Otto CM, Rieser TM, Brooks MB, Russell MW. Evidence of hypercoagulability in dogs with parvoviral enteritis.. J Am Vet Med Assoc 2000;217(10):1500‐1504.
    doi: 10.2460/javma.2000.217.1500pubmed: 11128540google scholar: lookup
  31. Boutet P, Heath F, Archer J, Villiers E. Comparison of quantitative immunoturbidimetric and semiquantitative latex‐agglutination assays for D‐dimer measurement in canine plasma. Vet Clin Path 2009;38(1):78‐82.
    doi: 10.1111/j.1939-165x.2008.00083.xpubmed: 19228362google scholar: lookup
  32. Caldin M, Furlanello T, Lubas G. Validation of an immunoturbidimetric D‐dimer assay in canine citrated plasma. Vet Clin Path 2000;29(2):51‐54.
    doi: 10.1111/j.1939-165x.2000.tb00398.xpubmed: 12070811google scholar: lookup
  33. Bauer N, Moritz A. Evaluation of the cardiac reader® as a point‐of‐care instrument for measurement of fibrin D‐dimers in dogs. Tierärztliche Praxis Ausgabe K Kleintiere Heimtiere 2009;37(5):319‐325.
    doi: 10.1055/s-0038-1622809google scholar: lookup
  34. Armengou L, Monreal L, Tarancón I, Navarro M, Ríos J, Segura D. Plasma D‐dimer concentration in sick newborn foals. J Vet Intern Med 2008;22(2):411‐417.
    doi: 10.1111/j.1939-1676.2008.0050.xpubmed: 18312557google scholar: lookup
  35. Stokol T, Erb HN, Wilde LD, Tornquist SJ, Brooks M. Evaluation of latex agglutination kits for detection of fibrin(ogen) degradation products and D‐dimer in healthy horses and horses with severe colic. Vet Clin Path 2005;34(4):375‐382.
    doi: 10.1111/j.1939-165x.2005.tb00064.xpubmed: 16270263google scholar: lookup
  36. Stokol T, Brooks M, Rush JE. Hypercoagulability in cats with cardiomyopathy. J Vet Intern Med 2008;22(3):546‐552.
    doi: 10.1111/j.1939-1676.2008.0098.xpubmed: 18466239google scholar: lookup
  37. Kogan AE, Mukharyamova KS, Bereznikova AV. Monoclonal antibodies with equal specificity to D‐dimer and high‐molecular‐weight fibrin degradation products. Blood Coagul Fibrinolysis 2016;27(5):542‐550.
    doi: 10.1097/mbc.0000000000000453pmc: PMC4935535pubmed: 26656897google scholar: lookup
  38. Russell EB, Courtman NF, Santos LL, Tennent‐Brown BS. Fibrinogen heterogeneity in horses. J Vet Intern Med 2021;35(2):1131‐1139.
    doi: 10.1111/jvim.16065pmc: PMC7995376pubmed: 33604912google scholar: lookup
  39. Dempfle CE, Zips S, Ergül H, Heene DL, FACT Study Group. The fibrin assay comparison trial (FACT): correlation of soluble fibrin assays with D‐dimer. Thromb Haemostasis 2001;86(5):1204‐1209.
    pubmed: 11816708
  40. Bennick A, Haddeland U, Brosstad F. D‐dimer specific monoclonal antibodies react with fibrinogen aggregates. Thromb Res 1996;82(2):169‐176.
    doi: 10.1016/0049-3848(96)00063-1pubmed: 9163070google scholar: lookup
  41. Miura N, Furukawa M, Magari Y, Momoi Y. Cross‐reactivity of the anti‐human D‐dimer monoclonal antibody 1C9‐6F10 to canine fibrin degradation products. J Vet Med Sci 2013;75(7):963‐966.
    doi: 10.1292/jvms.12-0270pubmed: 23420644google scholar: lookup
  42. Devine DV, Greeberg CS. 223 Monoclonal antibody to fibrin d‐dimer (DD‐3B6) recognizes an epitope on the gamma chain of fragment D. Fibrinolysis 1988;2:95.
    doi: 10.1016/0268-9499(88)90570-xpubmed: 2451885google scholar: lookup

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 100,367 horse owners like you who receive our email updates on horse health and nutrition.