Hereditary spherocytosis (HS) is a common inherited erythrocyte membrane disorder characterized by chronic hemolytic anemia. Clinical manifestations and biochemical abnormalities of HS are heterogeneous. In this study, we investigated erythrocyte membrane protein defects in 27 Korean HS cases. Utilizing both the Fairbanks system and the Laemmli system, sodium dodecyl sulfate polyacrylamide gel electrophoresis of erythrocyte membrane proteins was performed. Proteins were stained with Coomassie brilliant blue and gels were scanned using a densitometer. We detected spectrin deficiency in 7.4% of cases (2/27), ankyrin deficiency in 29.6% (8/27), combined spectrin and ankyrin deficiency in 3.7% (1/27), band 3 deficiency in 11.1% (3/27) and protein 4.2 deficiency in 14.8% (4/27). Membrane protein deficiencies were not observed in nine cases (33.3%, 9/27). Members of two of seven families tested showed the same protein defects as the proband. Ankyrin deficiency alone and combined with spectrin deficiency accounted for 33.3% of cases (9/27), and they were the most common biochemical defects in Korean HS cases. Protein 4.2 deficiency caused HS more frequently in Koreans than in Caucasians.
Ankyrins/analysis* ; Band 3 Protein/analysis* ; Erythrocyte Membrane/chemistry* ; Human ; Korea ; Spectrin/analysis* ; Spherocytosis, Hereditary/blood*

Hereditary spherocytosis (HS) is a common inherited erythrocyte membrane disorder characterized by chronic hemolytic anemia. Clinical manifestations and biochemical abnormalities of HS are heterogeneous. In this study, we investigated erythrocyte membrane protein defects in 27 Korean HS cases. Utilizing both the Fairbanks system and the Laemmli system, sodium dodecyl sulfate polyacrylamide gel electrophoresis of erythrocyte membrane proteins was performed. Proteins were stained with Coomassie brilliant blue and gels were scanned using a densitometer. We detected spectrin deficiency in 7.4% of cases (2/27), ankyrin deficiency in 29.6% (8/27), combined spectrin and ankyrin deficiency in 3.7% (1/27), band 3 deficiency in 11.1% (3/27) and protein 4.2 deficiency in 14.8% (4/27). Membrane protein deficiencies were not observed in nine cases (33.3%, 9/27). Members of two of seven families tested showed the same protein defects as the proband. Ankyrin deficiency alone and combined with spectrin deficiency accounted for 33.3% of cases (9/27), and they were the most common biochemical defects in Korean HS cases. Protein 4.2 deficiency caused HS more frequently in Koreans than in Caucasians.
Ankyrins/analysis* ; Band 3 Protein/analysis* ; Erythrocyte Membrane/chemistry* ; Human ; Korea ; Spectrin/analysis* ; Spherocytosis, Hereditary/blood*

OBJECTIVETo investigate the sensitivity and specificity of eosin-5'-maleimide (EMA)assay for the diagnosis of hereditary spherocytosis (HS), and to verify the stability of reagent and samples.

METHODSEMA flow cytometry test, NaCl-osmotic fragility test and acidified glycerol lysis test were performed using peripheral blood samples from 80 patients with HS and 44 patients with other blood diseases, the sensitivity and specificity of the three methods were compared, and the feasibility of EMA binding test was estimated. The stability of EMA reagent and HS samples stored at different temperatures were tested.

RESULTSAmong the 124 tested samples, the sensitivity and specificity of EMA binding test was 0.925 and 0.954, that of NaCl-osmotic fragility test was 0.950 and 0.455, and that of acidified glycerol lysis test was 1.000 and 0.318, respectively. Although the sensitivity of NaCl-osmotic fragility test and acidified glycerol lysis test was a little higher than that of EMA binding test, the specificity of the former two methods was poor, they couldn't clearly distinguish whether spherocytosis is hereditary spherocytosis. The experiment results showed that EMA was sensitive to the temperature and should not be stored in a small aliquots at -80 ℃ over a period of 6 months. The stability of the HS sample was better, 6 days storage at 4 ℃ and 3 days storage at room temperature had no influence on the results.

CONCLUSIONEMA binding test by flow cytometry showed good sensitivity and specificity for HS diagnosis. EMA reagent should be stored at-80 ℃ and the HS samples should be tested within 6 days storage at 4 ℃ and 3 days at room temperature.

Ankyrins ; blood ; deficiency ; Eosine Yellowish-(YS) ; analogs & derivatives ; Flow Cytometry ; Hematologic Tests ; Humans ; Sensitivity and Specificity ; Spherocytosis, Hereditary ; blood ; diagnosis

INTRODUCTIONThe underlying diagnosis of severe anaemic illnesses in children may not be easy to identify at times, especially when regular blood transfusion has been started.

MATERIALS AND METHODSInternational children patients attending a haematology clinic for diagnostic evaluation were identified retrospectively if they had to receive repeated blood transfusions with an undiagnosed illness or an incorrect diagnosis. Their demographic data, presenting features, and eventual diagnosis were described.

RESULTSTwelve children including 7 boys were enrolled from March 2007 to August 2011. Five came from Vietnam; 2 each came from Bangladesh and Indonesia; and 1 each from Hong Kong, Myanmar, and Ukraine. Their illnesses started at a mean age of 1.5 years (0.1 to 6.6) and they had been receiving blood transfusion for a mean duration of 2.5 years (0.1 to 9.9) years prior to the evaluation. Thalassemia major was the fi rst diagnosis in 5 cases; one had been treated for autoimmune haemolytic anaemia while the rest had not been given a diagnosis. After the evaluation, 4 children were diagnosed with Diamond Blackfan anaemia, 3 were diagnosed with hereditary spherocytosis, and one each with hereditary pyropoikilocytosis, congenital sideroblastic anaemia, congenital thrombotic thrombocytopenic purpura, transient erythroblastopenia of childhood, and autoimmune myelofibrosis associated with human immunodeficiency virus infection.

CONCLUSIONA definitive diagnosis can be identified in this cohort of children on medical tourism with severe anaemic illnesses requiring repeated transfusions with diagnostic approaches that circumvent the interference of transfused cells.

Anemia ; diagnosis ; therapy ; Anemia, Diamond-Blackfan ; diagnosis ; Blood Transfusion ; Child ; Child Health Services ; Child, Preschool ; Delayed Diagnosis ; Diagnostic Errors ; Female ; Humans ; Infant ; Male ; Medical Tourism ; Retrospective Studies ; Spherocytosis, Hereditary ; diagnosis