Determination of erythrocyte membrane proteins in children with the diagnosis of hereditary spherocytosis by ELISA

Objective: Hereditary spherocytosis (HS) is one of the most common causes of hereditary hemolytic anemia among erythrocyte membrane disorders. Because of the important clinical, laboratory, biochemical and genetic diversity in the patients, it is important to make a correct diagnosis. In our study, we aimed to determine the erythrocyte membrane proteins leading to HS by ELISA. Materials and Methods: Between the years 2004-2017 in the Pediatric Hematology Department of Adnan Menderes University Medical School, all HS patients were included in the study, which is considered HS with history, physical examination and laboratory findings and diagnosed with positivity osmotic fragility test. Ankyrin, spectrin α, spectrin β, band 3 and protein 4,2 were studied by ELISA. Statistical analyzes were performed via SPSS packet program. Kolmogorov-Smirnov test, T test and discriminant analysis were used in the analyses. P <0.05 was regarded as statistically significant. Results: Twenty HS patients and 20 control patients were included in the study. In 30% of patients, the ankyrin level; in 10% of patients, the spectrin α level; in 25% of patients, spectrin β level; in 10% of patients, band 3 level; in 45% of patients protein 4,2 level were found to be lower than in control group. In 35% of patients was found lack of combined protein with at least two protein deficiencies. 71,4% of the patients with combined protein deficiency were severe, 14,3% were moderate, 14,3% were mild HS patients and 71.4% were female. There was no statistically significant difference between ankyrin and band 3 levels between patient and control group. Spectrin α and protein 4,2 levels were significantly lower in the patient group when compared with the control group. The most significant difference was found in the spectrin β level. Of all the proteins, the lowest in the patient group and the most statistically significant were the spectrin β. The sensitivity, specificity, positive and negative predictive value of the ELISA test were 75% in setting HS diagnosis. Conclusion: In this study, spectrin α, spectrin β and protein 4,2 levels were found to be important variables in discriminating patients and control group. It was observed that the combined erythrocyte membrane protein deficiency may cause severe HS clinic. Therefore, we think that it is appropriate to investigate the presence of combined protein deficiency in such patients. In addition to the presence of six normal patients with erythrocyte membrane proteins, a low number of patients suggests that higher sensitivity and specificity rates may not be achieved. It also revealed that qualitative protein impairment may be detected in these patients whose quantitative membrane proteins are normal, and it is necessary to evaluate them in this respect. The data we have obtained show that ELISA can be preferred in research because of its sensitivity and practicality and quantitative results. However, there is a need for more extensive work to be done on this issue. Keywords: Hereditary spherocytosis, erythrocyte membrane, ankyrin, spectrin, band 3, protein 4,2, ELISA, child