Rapid flow cytometric test for the diagnosis of membrane cytoskeleton-associated haemolytic anaemia

Br J Haematol. 2000 Dec;111(3):924-33.


The flow cytometric test measures the fluorescence intensity of intact red cells labelled with the dye eosin-5-maleimide, which reacts covalently with Lys-430 on the first extracellular loop of band 3 protein. In this study, red cells from patients with hereditary spherocytosis (HS), congenital dyserythropoietic anaemia type II, South-east Asian ovalocytosis and cryohydrocytosis have produced a greater degree of reduction of mean channel fluorescence readings than those for other patient groups and normal controls. The predictive value of this test for membrane abnormality was compared with the results obtained from the sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) method, which is currently the reference laboratory test for the identification of membrane protein deficiencies in hereditary spherocytosis and for the detection of spectrin variants in hereditary elliptocytosis. The dye method is a reliable, speedy diagnostic test (2 h from sample collection to result) for HS with a sensitivity of 92.7% and a specificity of 99.1%. Thus, it will serve well as a first-line screening test for the diagnosis of hereditary spherocytosis in routine haematology.

Publication types

  • Comparative Study

MeSH terms

  • Adult
  • Anemia, Hemolytic, Congenital / diagnosis*
  • Anion Exchange Protein 1, Erythrocyte / metabolism
  • Chymotrypsin / metabolism
  • Cytoskeleton / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Eosine Yellowish-(YS) / analogs & derivatives*
  • Eosine Yellowish-(YS) / metabolism
  • Erythrocyte Membrane / metabolism
  • Fetal Blood / cytology
  • Flow Cytometry / methods
  • Fluorescent Dyes / metabolism
  • Humans
  • Infant, Newborn
  • Predictive Value of Tests
  • ROC Curve
  • Spherocytosis, Hereditary / diagnosis


  • Anion Exchange Protein 1, Erythrocyte
  • Fluorescent Dyes
  • eosin maleimide
  • Chymotrypsin
  • Eosine Yellowish-(YS)