以流式細胞儀建立診斷遺傳性球型紅血球增多症之標準檢驗方法

Flow Cytometry as a Standard Procedure for the Diagnosis of Hereditary Spherocytosis

吳佳蓉(Jia-Rong Wu) ； 施佩君(Pei-Chun Shih) ； 周文堅(Wen-Chien Chou) ； 許呈安(Cheng-An Hsu)

《Journal of Biomedical & Laboratory Sciences》 29卷2期 (2017/06) Pp. 49-54

摘要

遺傳性球型紅血球增多症(HS)是一種紅血球膜結構上的遺傳性異常，傳統上以紅血球脆性試驗進行篩檢，此項試驗非常的耗時、費力，且至少須取得9 ml的全血。EMA是一種螢光染劑，會與紅血球膜蛋白band 3上的Lys-430形成共價性鍵結；HS病患band 3蛋白數量會有下降的狀況，造成EMA螢光強度下降。為增進HS病患的篩檢能力，我們標準化流式細胞儀偵測EMA的步驟，透過ROC curve取得正常人與病患的cut-off值，並且分析HS病患分別在EMA試驗與脆性試驗的結果表現。總結來說，與脆性試驗相較，EMA試驗擁有較佳的靈敏度及專一性，操作簡易，數小時內便可得到結果。EMA試驗僅需少量的全血檢體，也適合在新生兒需要篩檢時使用。

關鍵字

遺傳性球型紅血球增多症 ； Eosin-5-maleimide ； EMA

並列摘要

Hereditary spherocytosis (HS) is a hereditary disorder of membrane structure on red blood cells. Traditional laboratory screening for HS is osmotic fragility test, which is labor-intensive and time-consuming requiring at least 9 ml of whole blood. EMA is a fluorescence dye, which binds covalently Lys-430 on the band 3 RBC membrane protein. The decrease of band 3 protein in HS patient results in the lower EMA fluorescence intensity. To improve the HS screening, we standardize the protocol of EMA flow cytometry. Set up a cut-off value through ROC curve to discriminate the normal individuals and HS patients. We also compare the performance between the EMA flow cytometry and OF test in a series of HS patients. Compared to the OF test, the EMA test has a better sensitivity and specificity, is easier to practice, and the results are available in hours. The EMA test needs small volume of whole blood, more suitable for neonatal screening.

並列關鍵字

Hereditary spherocytosis ； Eosin-5-maleimide ； EMA