用于RhD抗原流式细胞检测的固定透化方法评价

发布时间：2018-01-12 02:28

  本文关键词：用于RhD抗原流式细胞检测的固定透化方法评价　出处：《中国输血杂志》2017年09期 　论文类型：期刊论文

  更多相关文章： 甲醛 多聚甲醛 戊二醛 固定 透化 RhD抗原

【摘要】：目的寻找具有回收率高且适于RhD抗原荧光检测的红细胞固定透化方法。方法分别采用4%甲醛、4%多聚甲醛、2%戊二醛对红细胞进行固定,同时采用4%甲醛-Triton X-100、4%多聚甲醛-Triton X-100、2%戊二醛-Triton X-100对红细胞进行透化固定,未处理红细胞作为对照。冰上固定20 min,洗涤后计算红细胞回收率。在各组红细胞中加入1∶128的RhD Ig G单克隆抗体,同时设组内空白对照,37℃孵育30 min。PBS洗涤3次后加入1∶200的FITC荧光抗体,37℃孵育30 min,PBS洗涤3次后流式细胞仪检测。结果 2%戊二醛及2%戊二醛-Triton X-100 2组的回收率分别为72.83%与72.80%,高于对照组(58.32%),甲醛及多聚甲醛2组的回收率在4.00%左右,远低于对照组。流式检测结果显示,不同固定剂的荧光本底均高于未处理红细胞。与对照组相比甲醛与多聚甲醛会减弱RhD抗原荧光强度,而戊二醛不影响RhD荧光强度,戊二醛-Triton X-100可增强RhD抗原检测的灵敏度。结论2%戊二醛-Triton X-100固定透化法不仅可得到较高的红细胞回收率,且RhD荧光检测的灵敏度也得到提升,是荧光检测RhD抗原较理想的固定透化方法。
[Abstract]:Objective to find a method of erythrocyte fixation and permeation with high recovery and suitable for RhD antigen fluorescence detection. Methods 4% formaldehyde 4% paraformaldehyde 2% glutaraldehyde was used to immobilize erythrocyte. At the same time, 4% formaldehyde -Triton X-100 was used. 2% glutaraldehyde Triton X-100 was used to immobilize red blood cells, and untreated red blood cells were used as control, and fixed on ice for 20 min. The recovery rate of red blood cells was calculated after washing. 1: 128 monoclonal antibody of RhD Ig G was added to the erythrocytes of each group, and a blank control group was set up at the same time. Incubated at 37 鈩，

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