Abstract
Hereditary spherocytosis (HS) is a common syndrome characterized by abnormal quantities of red cell membrane proteins. In this study, we attempted to establish a simple diagnostic technique that can discriminate between HS and other conditions, such as congenital hemolytic anemia, β thalassemia (β thal), hemoglobinopathy (HP), autoimmune hemolytic anemia (AIHA), erythroenzymopathy, and iron deficiency anemia. We studied peripheral blood red cells stained with standard Wright–Giemsa and eosin-5′-maleimide (EMA), and evaluated red cell hemoglobin peak patterns by high-performance liquid chromatography (HPLC). Furthermore, the remainder of the blood sample was biochemically analyzed for quantities of red cell membrane proteins by SDS-polyacrylamide gel electrophoresis (PAGE). Red blood cells from pre-splenectomized HS (pre-HS) patients were faintly stained with EMA, whereas those from other patients and controls were brilliantly stained. Pre-HS patients exhibited lower quantities of the band 3 protein than patients with other anemias. The hemoglobin peak patterns showed abnormal and characteristic waves for patients with HP, β thal, and AIHA, but not in pre-HS or post-splenectomized HS (post-HS) patients, IDD, and controls. Pre-HS patients with low quantities of the band 3 protein showed red blood cells faintly stained with EMA, which correlated with HS and other hemolytic anemias. Red blood cells from post-HS patients were stained similarly to those from controls but were stained more brilliantly than those from pre-HS patients. Thus, these two simple techniques, EMA staining and HPLC, along with SDS-PAGE, enable the discrimination between the initial stages of HS and other anemias. However, DNA analyses may be necessary for cases where these methods cannot adequately distinguish between the conditions.
