本文關(guān)鍵詞：用于RhD抗原流式細(xì)胞檢測的固定透化方法評價(jià)　出處：《中國輸血雜志》2017年09期 　論文類型：期刊論文

  更多相關(guān)文章： 甲醛 多聚甲醛 戊二醛 固定 透化 RhD抗原

【摘要】：目的尋找具有回收率高且適于RhD抗原熒光檢測的紅細(xì)胞固定透化方法。方法分別采用4%甲醛、4%多聚甲醛、2%戊二醛對紅細(xì)胞進(jìn)行固定,同時(shí)采用4%甲醛-Triton X-100、4%多聚甲醛-Triton X-100、2%戊二醛-Triton X-100對紅細(xì)胞進(jìn)行透化固定,未處理紅細(xì)胞作為對照。冰上固定20 min,洗滌后計(jì)算紅細(xì)胞回收率。在各組紅細(xì)胞中加入1∶128的RhD Ig G單克隆抗體,同時(shí)設(shè)組內(nèi)空白對照,37℃孵育30 min。PBS洗滌3次后加入1∶200的FITC熒光抗體,37℃孵育30 min,PBS洗滌3次后流式細(xì)胞儀檢測。結(jié)果 2%戊二醛及2%戊二醛-Triton X-100 2組的回收率分別為72.83%與72.80%,高于對照組(58.32%),甲醛及多聚甲醛2組的回收率在4.00%左右,遠(yuǎn)低于對照組。流式檢測結(jié)果顯示,不同固定劑的熒光本底均高于未處理紅細(xì)胞。與對照組相比甲醛與多聚甲醛會減弱RhD抗原熒光強(qiáng)度,而戊二醛不影響RhD熒光強(qiáng)度,戊二醛-Triton X-100可增強(qiáng)RhD抗原檢測的靈敏度。結(jié)論2%戊二醛-Triton X-100固定透化法不僅可得到較高的紅細(xì)胞回收率,且RhD熒光檢測的靈敏度也得到提升,是熒光檢測RhD抗原較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，

本文編號：1412310