Diagnosis and molecular analysis of red cell enzyme anomalies and analysis of red cell deformabilities

点击翻译按钮获取中文摘要

We discovered 16 cases of glucose-6-phosphate dehydrogenase (G6PD) deficiency, 7 cases of pyruvate kinase (PK) deficiency, a case of phosphofructokinase deficiency, and a case of phosphoglycerate kinase deficiency by the enzymatic analysis of 117 cases associated with hereditary non-spherocytic hemolytic anemia.PK deficiency is the most common erythroenzymopathy associated with hereditary non-spherocytic hemolytic anemia due to a glycolytic enzyme defect. We determined three mutations (664-6 ins GAC, 1468T, 1436A) by the analysis of 5 Japanese PK-deficient families. In addition, we elucidated the new homozygous mutation (1231A) among a Korean boy, which is the first Korean PK deficiency determined the moleculer abnormalities at the gene level.G6PD A is a common G6PD variant among Africans that may cause acute hemolysis triggered by infections and certain drugs, as well as by fava beans. This phenotype can be caused by a combination of the common 376G mutation and either of 3 additional mutations: 202A, 680T, or 968T. We discovered a Japanese boy of G6PD deficiency associated with acute hemolysis. Using PCR-SSCP analysis combined with direct sequencing, we identified a missense mutation 202A, but failed to find the counterpart 376G mutation or any additional mutation. We concluded that a single mutation 202A in the human G6PD gene can cause acute hemolysis by itself.We examined the red cell deformabilities between normal and PK-deficient conditions at a single cell level using the microchannel array. We could not obtain the microchannel less than 6 μm. Therefore, it was impossible to demonstrate the apparent result of the reduced red cell deformabilities under the ATP-deficient state at the physiological condition.