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  • Title: [Flow cytometric test using eosin-5'-maleimide (EMA) labelling of red blood for diagnosis of hereditary spherocytosis].
    Author: Wang J, Zheng B, Zhao Y, Chen X, Liu Y, Bo L, Zheng Y, Zhang F, Ru K, Wang H.
    Journal: Zhonghua Xue Ye Xue Za Zhi; 2015 Jul; 36(7):598-601. PubMed ID: 26304086.
    Abstract:
    OBJECTIVE: To investigate the sensitivity and specificity of eosin-5'-maleimide (EMA)assay for the diagnosis of hereditary spherocytosis (HS), and to verify the stability of reagent and samples. METHODS: EMA flow cytometry test, NaCl-osmotic fragility test and acidified glycerol lysis test were performed using peripheral blood samples from 80 patients with HS and 44 patients with other blood diseases, the sensitivity and specificity of the three methods were compared, and the feasibility of EMA binding test was estimated. The stability of EMA reagent and HS samples stored at different temperatures were tested. RESULTS: Among the 124 tested samples, the sensitivity and specificity of EMA binding test was 0.925 and 0.954, that of NaCl-osmotic fragility test was 0.950 and 0.455, and that of acidified glycerol lysis test was 1.000 and 0.318, respectively. Although the sensitivity of NaCl-osmotic fragility test and acidified glycerol lysis test was a little higher than that of EMA binding test, the specificity of the former two methods was poor, they couldn't clearly distinguish whether spherocytosis is hereditary spherocytosis. The experiment results showed that EMA was sensitive to the temperature and should not be stored in a small aliquots at -80 ℃ over a period of 6 months. The stability of the HS sample was better, 6 days storage at 4 ℃ and 3 days storage at room temperature had no influence on the results. CONCLUSION: EMA binding test by flow cytometry showed good sensitivity and specificity for HS diagnosis. EMA reagent should be stored at-80 ℃ and the HS samples should be tested within 6 days storage at 4 ℃ and 3 days at room temperature. 目的: 探讨伊红-5′-马来酰亚胺（EMA）为标记流式细胞术检测遗传性球形红细胞的灵敏性和特异性，并对试剂和样本稳定性进行验证。 方法: 对80例遗传性球形红细胞增多症（HS）和44例非HS患者外周血样本全部采用EMA标记流式细胞术检测、红细胞渗透脆性试验和酸化甘油溶解试验三种方法进行检测，比较三种方法的灵敏性和特异性，探讨EMA标记流式细胞术检测的可行性。并观察EMA及样本在不同储存条件下的稳定性。 结果: 通过检测124例样本得出，EMA标记流式细胞术检测的灵敏性和特异性分别为0.925和0.954，红细胞渗透脆性试验分别为0.950和0.455，酸化甘油溶解试验为1.000和0.318。红细胞渗透脆性试验和酸化甘油溶解试验的灵敏性稍高于EMA流式检测方法，但特异性差，不能明确区分球形红细胞增多是否为HS。EMA对温度很敏感，−80 °C储存180 d较4 °C存储1 d稳定。HS样本的稳定性较好，4 °C放置6 d、室温条件放置3 d不会影响结果的判定。 结论: EMA标记流式细胞术检测HS具有良好的灵敏性和特异性，−80 °C储存EMA较为稳定，须在样本4 °C放置6 d内、室温条件放置3 d内完成检测。
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