流式细胞术检测红细胞渗透脆性试验方法的建立

Establishment of Method for Detecting Red Blood Cell Osmotic Fragility by Flow Cytometry

目的:建立流式细胞术检测红细胞渗透脆性的方法。方法:将红细胞悬浮于由Nacl液和去离子水制备的不同浓度低渗盐溶液(0.70 ml生理盐水+0.3 ml去离子水,0.60 ml生理盐水+0.40 ml去离子水和0.55 ml生理盐水+0.45 ml去离子水)中,利用流式细胞仪及时测定残留红细胞数量。结果:3种NaCl稀释浓度中,流式细胞仪检测的不同时间区域剩余红细胞数量百分比差别不大。选取B+C+D+E+F+G区域比较,不同NaCl稀释浓度下3组的剩余红细胞数量百分比均有差异;正常对照组的剩余红细胞百分比显著低于血红蛋白病组;遗传性球形红细胞增多症(hereditary spherocytosis,HS)组显著低于血红蛋白病组和正常组(P<0.05)。3种不同稀释浓度比较发现第2种浓度(0.60 ml N.S.+0.40 ml H_2O)更适合于本实验室Beckman-Coulter流式细胞仪FC500对遗传性球形红细胞增多症的筛查。结论:选用流式细胞术检测红细胞渗透脆性简单、快速、客观、经济。本方法可能是一种非常有效的对遗传性球形红细胞进行初筛的方法。

Objective:To establish a new method for detection of red blood cell osmotic fragility by using flow cytometry.Methods:The hypotension salt solution of different concentrations(0.70 ml normal saline + 0.3 ml deionized water,0.60 ml normal saline +0.40 ml deionized water and 0.55 ml normal saline +0.45 ml deionized water) were prepared with normal saline and deionized water,in which the red blood cells were suspended,and the residual red blood cells were detected by flow cytometer.Results:There was no significant difference in percentage of residual red blood cells between different time points detected by flow cytometer in 3 different hypotonic salt solutions.The percentage of residual red blood cells in B+C+D+E + F + G detected time region was different among 3 NaCl dilution groups.The percentage of residual red blood cells in normal control was lower than that in hemoglobinopathy group.The percentage of residual red blood cells in hereditary spherocytosis(HS) group was obviously lower than that in hemoglobinopathy and normal control groups.The comparison of 3 different dilution concentrations found that the second concentration(0.60 ml normal saline +0.40 ml deionized water) is more suitable to screen HS by FC500 flow cytometer.Conclusion:The detection of red cell osmotic fragility by using flow cytometry is a simple,rapid,objective and economic way that can be an effective screening method for diagnose the HS.