Evaluation of eosin-5-maleimide flow cytometric test in diagnosis of hereditary spherocytosis

• Published in: International journal of laboratory hematology 2010-02, Vol.32 (1p2), p.8-16
• Main Authors: KAR, R., MISHRA, P., PATI, H. P.
• Format: Article
• Language: English
• Subjects: Anemia, Hemolytic - diagnosis; Diagnosis, Differential; eosin-5-maleimide; Eosine Yellowish-(YS) - analogs & derivatives; Eosine Yellowish-(YS) - analysis; Flow cytometry; Flow Cytometry - methods; fluorescence; hereditary spherocytosis; Humans; Reference Standards; Sensitivity and Specificity; Spherocytosis, Hereditary - diagnosis
• ISSN: 1751-5521; 1751-553X
• DOI: 10.1111/j.1751-553X.2008.01098.x

Summary A flow cytometry‐based test using eosin‐5‐maleimide (EMA) dye was used for diagnosis of hereditary spherocytosis (HS). The mean fluorescence intensiy (MFI) of EMA tagged erythrocytes is lower in HS than that in other hemolytic and nonhemolytic anemias. We enrolled 114 subjects comprising 20 confirmed HS, 20 suspected HS/hemolytic anemia (HA), 20 normal controls, 20 other hemolytic anemias [13 autoimmune hemolytic anemia, three congenital dyserythropoietic anemia (CDA), one pyruvate kinase deficiency, two microangiopathic hemolytic anemia], 18 microcytic anemia and 16 macrocytic anemia cases. All samples were subjected to flow cytometry as per standard protocol. The mean MFI of normal control subjects was 11 861.5 (SD 883.5) and of confirmed HS was 7949.3 (SD 1304.1). Using this test, of 20 patients suspected to be HS/HA but with no confirmatory diagnosis, eight patients were diagnosed as HS. Using logistic regression analysis, the optimum cut‐off MFI value between HS and normal controls was 10126. The area under the ROC curve was 0.99. The statistical significance of MFI values was obtained by t‐test or Wilcoxon rank sum test as applicable. Compared with normal controls, the MFI values in HS were lower and in megaloblastic anemia were higher which was statistically highly significant (P