Rapid flow cytometric test for the diagnosis of hereditary spherocytosis and its relevance to clinical severity

ABSTRACT

The causes of non-immunehereditary hemolyticanaemia include enzymopathy, hemoglobinopathy, abnormal cation transport and membranopathy. Hereditary spherocytoris [HS] is the most common membranopathy. In this study a new flow cytometic test measures the fluorescence in tensity of intact red cells labeled with eosin -5- maliernide. The dye reacts covalentily with lys-430 on the first extracellular loop of band 3 protein. In this study, red cells from patients with HS produced a greater degree of reduction of mean channel fluorescence [MCF] readings than those for other patients groups with other causes of anaemia and normal controls. The MCF for spherocytic patients was 20.4 +/- 5.2 as compared to the control group whose MCF was 35.9 +/- 2.6. The MCF readings of other patients groups included in the study were for thalassaemia patient [39.8 +/- 2.2], glucose -6- phosphate dehydrogenase deficiency patients [34.8 +/- 1.9] iron deficiency anaemia patient [30.25 +/- 1.2], autoimmune hemolytic anaemia patients [32.5 +/- 1.7] and chronic renal failure patients [34.6 +/- 1.7]. All these groups of patients gave MCF readings that were higher than those of HS. Thus the dye method proved to be a reliable and speedy diagnostic test [2 hours from sample collection to result] for the diagnosis of hereditary spherocytosis in routine hematology