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  • Title: Rapid flow cytometric test using eosin-5-maleimide for diagnosis of red blood cell membrane disorders.
    Author: Tachavanich K, Tanphaichitr VS, Utto W, Viprakasit V.
    Journal: Southeast Asian J Trop Med Public Health; 2009 May; 40(3):570-5. PubMed ID: 19842445.
    Abstract:
    Conventional diagnosis of hereditary red blood cell (RBC) membrane disorders, in particular hereditary spherocytosis (HS), is labor intensive, time consuming and requires at least 2 ml of blood, which might be impractical in neonatal period. We evaluated the use of eosin-5-maleimide (EMA), a dye that reacts covalently with lysine-430 on the first extracellular loop of band 3 protein, for rapid screening test of patients with HS and Southeast Asian Ovalocytosis (SAO). Fresh RBCs from 142 healthy controls, 50 HS, 17 SAO, 29 hereditary elliptocytosis, 5 autoimmune hemolytic anemia, 66 patients with beta-thalassemia/HbE, 31 cases with alpha-thalassemia (HbH disease) and 4 cases with pyruvate kinase deficiency were stained with EMA, and analyzed for their mean channel fluorescence (MCF) using a flow cytometer. RBCs from patients with HS and SAO expressed a greater degree of reduction in MCF compared to those from normal controls and other hemolytic diseases. These findings showed that the fluorescence flow cytometric-based method is a simple, sensitive and reliable diagnostic test for RBC membrane disorders using a small volume of blood, and results could be obtained within 2 hours. Such method could serve as a first line screening for the diagnosis of HS and SAO in routine hematology before further specific membrane protein electrophoresis and molecular diagnosis are employed.
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