title: A comparative evaluation of Eosin-5'-maleimide flow cytometry reveals a high diagnostic efficacy for hereditary spherocytosis.
abstract: BACKGROUND
  Laboratory diagnosis of hereditary spherocytosis (HS) relies on increased incubated red cell osmotic fragility test for screening. We evaluated the diagnostic role of eosin-5'-maleimide (EMA) binding test by flow cytometry in spherocytic and microcytic hypochromic hematological disorders in North Indians.
  
  
  METHODS
  EMA binding test using flow cytometry was performed on 55 HS (40 families), 26 iron deficiency anemia (IDA), 32 β-thalassemia trait (βTT), and 10 autoimmune hemolytic anemia (AIHA) cases and 121 normals. Mean channel fluorescence (MCF) and coefficient of variation (CV) were studied. Different MCF parameters (MCF, MCF ratio, percent decrease MCF) and percent increase in CV were analyzed. Receiver operating characteristics analysis was performed to determine best cutoff values, sensitivity, and specificity for discriminating HS from other red cell disorders.
  
  
  RESULTS
  MCF ratio of HS group was significantly lower than normals (0.67 ± 0.07 vs. 1.01 ± 0.05, P < 0.001) and other cases. All patients with HS showed MCF ratio to be ≤0.79. Four postsplenectomy cases with near-normal hemograms also revealed low MCF ratio, showing the specificity of the test.
  
  
  CONCLUSIONS
  EMA assay was efficient to diagnose cases of HS including postsplenectomy cases and shows no overlap with IDA, βTT, and AIHA.
authors.affiliation: Department of Hematology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: P
authors.initials: P
authors.lastname: Joshi
authors.affiliation: Department of Hematology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: A
authors.initials: A
authors.lastname: Aggarwal
authors.affiliation: Department of Hematology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: M
authors.initials: M
authors.lastname: Jamwal
authors.affiliation: Department of Hematology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: M U S
authors.initials: MU
authors.lastname: Sachdeva
authors.affiliation: Department of Pediatrics, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: D
authors.initials: D
authors.lastname: Bansal
authors.affiliation: Department of Internal Medicine, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: P
authors.initials: P
authors.lastname: Malhotra
authors.affiliation: Department of Hematology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
authors.forename: P
authors.initials: P
authors.lastname: Sharma
authors.affiliation: Department of Hematology, Postgraduate Institute of Medical Education and Research, Chandigarh, India. das.reena@pgimer.edu.in, reenadaspgi@hotmail.com.
authors.forename: R
authors.initials: R
authors.lastname: Das
mesh_terms: D000293:Adolescent; D000328:Adult; D018798:Anemia, Iron-Deficiency / Q000097:blood / Q000175:diagnosis; D002648:Child; D002675:Child, Preschool; D004801:Eosine Yellowish-(YS) / Q000031:analogs & derivatives* / Q000494:pharmacology; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007223:Infant; D008297:Male; D008875:Middle Aged; D013103:Spherocytosis, Hereditary / Q000097:blood* / Q000175:diagnosis*; D017086:beta-Thalassemia / Q000097:blood / Q000175:diagnosis
chemical_list: C033531:eosin maleimide; D004801:Eosine Yellowish-(YS)
keywords: Eosin-5′-maleimide; North India; flow cytometer; hereditary spherocytosis