New Technology Capabilities of Direct Antiglobulin Test

EA Poponina, EV Butina, AV Iovdii, OD Maksimov, GA Zaitseva, IV Paramonov

Kirov Research Institute of Hematology and Transfusiology, 72 Krasnoarmeiskaya str., Kirov, Russian Federation, 610027

For citation: Elena Aleksandrovna Poponina, MD, PhD, 72 Krasnoarmeiskaya str., Kirov, Russian Federation, 610027; Tel.: +7(8332)54-51-83; e-mail:

For correspondence: Poponina EA, Butina EV, Iovdii AV, et al. New Technology Capabilities of Direct Antiglobulin Test. Clinical oncohematology. 2020;13(4):426–9. (In Russ).

DOI: 10.21320/2500-2139-2020-13-4-426-429


Background. Direct antiglobulin test (DAT) is used to identify erythrocyte-fixed antibodies and complement components. Gel methods are applied to differentiate immunoglobulin class and subclass in positive DAT, which allows to study the nature of anemia and assess the risk of immune hemolysis.

Aim. To assess the rate of positive DAT in oncohematological patients, to determine class and subclass of erythrocyte-fixed immunoglobulins, and to evaluate their contribution in hemolytic complications.

Materials & Methods. In 393 oncohematological patients at the Kirov Research Institute of Hematology and Transfusiology differentiated DAT was studied using gel test with Bio-Rad (USA) testing sets.

Results. The rate of positive DAT in oncohematological patients varied for different diseases from 6.2 % to 25.2 %, in the total group it was 15.5 %. It accounted for 6.2 % in acute leukemias, 6.3 % in myelodysplastic syndrome, 10 % in chronic myeloid leukemia, 11.9 % in Hodgkin’s lymphoma, 15.4 % in chronic lymphocytic leukemia, 21 % in non- Hodgkin’s lymphoma, and 25.2 % in multiple myeloma. In multiple myeloma, acute leukemia, Hodgkin’s lymphoma, and chronic myeloid leukemia patients the positive test was associated with IgG subclasses 2 and 4. In chronic lymphocytic leukemia and non-Hodgkin’s lymphoma patients IgG1 subclass 1, IgM and C3c, C3d complement components were detected on erythrocyte surfaces. It was shown that IgG2/IgG4 detection was not accompanied by any clinical or laboratory signs of immune hemolysis, IgG1 was responsible for destruction of erythrocytes in 50 % of cases, whereas the detection of C3c, C3d complement components was associated with hemolytic manifestations in 100 % of cases.

Conclusion. Positive DAT should be interpreted in light of laboratory and clinical data. Differentiated test helps to predict hemolytic complications in oncohematological patients.

Keywords: direct antiglobulin test, immunoglobulin class and subclass, immune hemolysis.

Received: April 27, 2020

Accepted: August 20, 2020

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  1. Sokol R, Hewitt S, Stamps B. Autoimmune haemolysis: an 18-year study of 865 cases referred to a regional transfusion center. Br Med J. 1981;282(6281):2023–7. doi: 10.1136/bmj.282.6281.2023.

  2. Минеева Н.В., Кробинец И.И., Бодрова Н.Н., Богданова И.О. Применение прямого антиглобулинового теста для выявления аутоантител при анемиях различного генеза. Онкогематология. 2017;12(3):57–62. doi: 10.17650/1818-8346-2017-12-3-57-62. [Mineeva NV, Krobinets II, Bodrova NN, Bogdanova IO. The use of direct antiglobulin test to detect antibodies in patients with anemia of various origins. Oncohematology. 2017;12(3):57–62. doi: 10.17650/1818-8346-2017-12-3-57-62. (In Russ)]

  3. Michel M. Classification and therapeutic approaches in autoimmune hemolytic anemia: an update. Expert Rev Hematol. 2011;4(6):607–18. doi: 10.1586/ehm.11.60.

  4. Judd W, Butch S, Oberman H, et al. The evaluation of a positive direct antiglobulin test in pretransfusion testing. Transfusion. 1980;20(1):17–23. doi: 10.1046/j.1537-2995.1980.20180125036.x.

  5. Packman C. Hemolytic anemia due to warm autoantibodies. Blood Rev. 2008;22(1):17–31. doi: 10.1016/j.blre.2007.08.001.

  6. Go R, Winters J, Kay N. How I treat autoimmune hemolytic anemia. Blood. 2017;129(22):2971–9. doi: 10.1182/blood-2016-11-693689.

  7. Berentsen S. Role of Complement in Autoimmune Hemolytic Anemia. Transfus Med Hemother. 2015;42(5):303–10. doi: 10.1159/000438964.

  8. Crisp D, Pruzanski W. B-Cell neoplasms with homogeneous cold-reacting antibodies (cold agglutinins). Am J Med. 1982;72(6):915–22. doi: 10.1016/0002-9343(82)90852-x.

  9. Jager U, D’Sa S, Schorgenhofer C, et al. Inhibition of complement C1s improves severe hemolytic anemia in cold agglutinin disease: a first-in-human trial. Blood. 2019;133(9):893–901. doi: 10.1182/blood-2018-06-856930.

  10. Parker V, Tormey C. The Direct Antiglobulin Test: Indications, Interpretation, and Pitfalls. Arch Pathol Lab Med. 2017;141(2):305–10. doi: 10.5858/arpa.2015-0444-rs.

  11. Тураев Р.Г., Бельская Е.Е. Проба Кумбса в лабораторной диагностике иммунопатологических состояний. Трансфузиология. 2017;1(18):59–64. [Turaev RG, Belskaya EE. Direct Coombs test in laboratory diagnosis of immunopathological states. Transfuziologiya. 2017;1(18):59–64. (In Russ)]

  12. Ricci F, Tedeschi A, Vismara E, et al. Should a Positive Direct Antiglobulin Test Be Considered a Prognostic Predictor in Chronic Lymphocytic Leukemia? Clin Lymphoma Myel Leuk. 2013;13(4):441–6. doi: 10.1016/j.clml.2013.02.024.

  13. Zantek N, Koepsell S, Tharp D, Cohn C. The direct antiglobulin test: A critical step in the evaluation of hemolysis. Am J Hematol. 2012;87(7):707–9. doi: 10.1002/ajh.23218.

  14. Rottenberg Y, Yahalom V, Shinar E, et al. Blood Donors with a Positive Direct Antiglobulin Test Are at Increased Risk for Hematologic Malignancies and Cancer in General. Blood. 2007;110(11):2903. doi: 10.1182/blood.v110.11.2903.2903.

  15. Rottenberg Y, Yahalom V, Shinar E, et al. Blood donors with positive direct antiglobulin tests are at increased risk for cancer. Transfusion. 2009;49(5):838–42. doi: 10.1111/j.1537-2995.2008.02054.x.

  16. Hannon J. Management of Blood Donors and Blood Donations From Individuals Found to Have a Positive Direct Antiglobulin Test. Transfus Med Rev. 2012;26(2):142–52. doi: 10.1016/j.tmrv.2011.08.004.

  17. Garratty G. The James Blundell Award Lecture 2007: Do we really understand immune red cell destruction? Transfus Med. 2008;18(6):321–34. doi: 10.1111/j.1365-3148.2008.00891.x.