摘要
目的建立流式细胞术快速检测红细胞渗透脆性的方法,评估流式细胞仪测定红细胞渗透脆性的可能性。方法收集30例抗凝外周血,每例标本均一分为二,1份在采集2 h内检测(正常对照组),另1份经37℃孵育24 h(阳性对照组);另收集3例临床确诊的遗传性球形红细胞增多症患者作为疾病组。分别使用流式细胞术检测各组红细胞渗透脆性。通过流式细胞术测定红细胞残留百分比确定生理盐水和无菌水最佳体积比,确定乙二胺四乙酸(EDTA)和肝素2种抗凝剂对其的影响。另收集14例肝素抗凝标本,同时采用改良Sanford法和流式细胞术测定红细胞残留百分比,对比评价流式细胞术和改良Sanford法测定红细胞渗透脆性的特点。结果采用流式细胞术检测正常对照组的红细胞残留百分比为72.03%±5.68%,明显高于阳性对照组(46.47%±15.83%,P<0.05);疾病组红细胞残留百分比为5.76%±3.54%,明显低于正常对照组(P<0.001)。当生理盐水和无菌水体积比为1.2∶0.8时,正常对照组和阳性对照组红细胞残留百分比差异最大(P<0.05);与EDTA抗凝剂比较,肝素抗凝的外周血更适合于红细胞渗透脆性检测。流式细胞术与改良Sanford法测定红细胞渗透脆性有较好的一致性,但流式细胞术敏感性更高。结论流式细胞术测定红细胞渗透脆性具有操作简便、敏感性更好等特点,有实际应用的可行性。
Objective To establish a rapid determination of erythrocyte osmotic fragility by flow cytometry, and to evaluate the feasibility of determining erythrocyte osmotic fragility byflow cytometry.Methods A total of 30 anticoagulationperipheral blood specimens were collected,and each specimen was divided into 2 parts,one group collected within 2 h as normal control group and another group incubated at 37℃ for 24 h as positive control group. A total of 3 patients diagnosed as hereditary spherocytosis were enrolled as disease group. The erythrocyte osmotic fragility was determined byflow cytometry. The residue percentage of red blood cells was analyzed byflow cytometry. The optimal volume ratio of normal saline and sterile water was identified. The influence of ethylene diamine tetraacetic acid（EDTA） and heparin anticoagulants was evaluated. A total of 14 cases of heparin anticoagulation specimens were collected, and the red blood cell residue percentage was determined by improved Sanford andflow cytometry simultaneously.The characterization of determining erythrocyte osmotic fragility byflow cytometry and improved Sanford was evaluated. Results Byflow cytometry,red blood cell residue percentages were 72.03% ± 5.68% in normal control group and 46.47% ± 15.83% in positive control group（P〈0.05）. The disease group showed a significant reduction in red blood cell residue percentage（5.76% ± 3.54%） compared to that in normal control group（P〈0.001）. When the volume ratio of normal saline to sterile water was 1.2∶0.8,there was significant difference of red blood cell residue percentage between the 2 groups（P〈0.05）. Compared with EDTA anticoagulant,heparin anticoagulant was more suitable for the determination of erythrocyte osmotic fragility. There was a good consistency betweenflow cytometry and improved Sanford inthe determination of erythrocyte osmotic fragility,butflow cytometry had better sensitivity than improved Sanford.Conclusions Flow cytometry has the characteristics of operation simplic
出处
《检验医学》
CAS
2016年第10期911-916,共6页
Laboratory Medicine
