Shibuya Children's Clinic, Sakado City, Japan.
Department of Biotechnology, Tokyo Denki University, Saitama, Tokyo, Japan.
Hematology. 2021 Dec;26(1):827-834. doi: 10.1080/16078454.2021.1979184.
In this study, clinical and biochemical methods were utilized to predict the final diagnosis of hereditary spherocytosis (HS), correlate the diagnosis with splenectomy, and examine the usefulness of this approach.
We biochemically and cytochemically analysed erythrocyte membrane proteins before making a final HS diagnosis based on gene analysis to compare diagnostic approaches. The clinical features of six patients with various subtypes of HS and symptoms were observed by blood analysis using eosin-5'-maleimide staining, biochemical analysis using sodium dodecyl sulphate - polyacrylamide gel electrophoresis with western blotting, and mass spectrometry. Finally, diagnostic membrane gene analysis was performed.
Five of the six patients showed mild to moderate or severe anaemia, and the other patient was non-anaemic; all six patients showed faint eosin-5'-maleimide staining. In western blotting of erythrocyte membrane proteins, all six patients (three with β-spectrin, two with ankyrin, and one with anomalies) showed low-molecular-weight peptide fragments, which were confirmed by mass spectrometry in the region corresponding to the band 3 protein. The two patients with an ankyrin gene anomaly exhibited severe anaemia, and two patients with simultaneous , , and anomalies exhibited mild anaemia and hyperbilirubinemia.
We determined the relationship among clinical features, cytochemical parameters, and gene anomalies in six patients with newly diagnosed HS while referring to previously published cases.
These findings reveal a close relationship between clinical features and membrane characteristics in HS, which can facilitate diagnosis and inform treatment.
本研究采用临床和生化方法预测遗传性球形红细胞增多症（HS）的最终诊断，将诊断与脾切除术相关联，并检验该方法的实用性。
我们在基于基因分析做出最终 HS 诊断之前，对红细胞膜蛋白进行了生化和细胞化学分析，以比较不同的诊断方法。通过使用曙红 5'-马来酰亚胺染色对 6 例具有不同 HS 亚型和症状的患者进行血液分析，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结合 Western blot 进行生化分析以及使用质谱法进行质量分析，观察其临床特征。最后进行诊断性膜基因分析。
6 例患者中有 5 例表现为轻至中度或重度贫血，另 1 例患者无贫血；所有 6 例患者均表现出微弱的曙红 5'-马来酰亚胺染色。在红细胞膜蛋白的 Western blot 中，所有 6 例患者（3 例β- spectrin、2 例锚蛋白、1 例异常）均显示低分子量肽片段，这些片段通过质谱法在与带 3 蛋白相对应的区域得到证实。2 例存在锚蛋白基因突变的患者表现为严重贫血，2 例同时存在 、 、 和 异常的患者表现为轻度贫血和高胆红素血症。
我们参考已发表的病例，确定了 6 例新诊断的 HS 患者的临床特征、细胞化学参数和基因异常之间的关系。
这些发现揭示了 HS 中临床特征与膜特征之间的密切关系，有助于诊断和治疗决策。
