RESUMEN
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.
Asunto(s)
Eliptocitosis Hereditaria/diagnóstico , Eosina Amarillenta-(YS)/análogos & derivados , Esferocitosis Hereditaria/diagnóstico , Adolescente , Adulto , Anemia Hemolítica Autoinmune/sangre , Estudios de Casos y Controles , Niño , Preescolar , Femenino , Citometría de Flujo , Humanos , Lactante , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad , Adulto Joven , Talasemia alfa/sangre , Talasemia beta/sangreRESUMEN
In order to provide a reference range for hematological parameters and red blood cells indices in Thai children, we analyzed data from 395 healthy non-anemic Thai children age from 1-16 years old, who all had normal pattern of hemoglobin typing (Hb A and Hb A2 less than 3.5%). Hematological analysis was performed using an automated cell counter and the hemoglobin studies were carried out by electrophoresis and liquid chromatography. Owing to a high frequency of a thalassemia in Thailand, cases with MCV < 75 fL has been excluded from the study since these cases were likely to be heterozygotes for alpha0 thalassemia. These criterions were applied to select so-called 'normal' controls for our analysis. Relatively mild microcytosis and hypochromia were observed, in particular in the first three years of age, suggesting an intrinsic immature nature of erythropiesis in the children. Age-dependent differences in the reference values for white blood cell (WBC) count and differential and platelet count were observed. Herein the hematological data and red blood cell indices were summarized according to ages and these will be of clinically useful for the future reference.