实用McAb-A-E直接法检验T淋巴细胞亚群PBMC细胞自然形态保存研究

A study of T lymphocyte subsets PBMC cell preservation of the natural form using practical Mc Ab-A-E direct method

目的:为了使实用Mc Ab-A-E直接法(单克隆抗体SPA红细胞花环法)适应临床实验室T淋巴细胞亚群检验快速、简便、准确的要求,对其细胞悬液不同方法制备和细胞自然形态保存研究。方法:选取20例临床标本,用自然沉降法(不加任何物质)和淋巴细胞分离液梯度密度离心法分别制取单个核细胞悬液,同时用实用Mc Ab-A-E直接法检验CD3+、CD4+、CD8+、CD4+/CD8+以及非淋巴细胞。结果:统计结果显示:两组CD3+(t=-4.197;P=0.000)、CD4+(t=-5.177;P=0.000)、CD8+(t=-4.24;P=0.000)、CD4+/CD8+(t=-1.758;P=0.095)和非淋巴细胞(t=-5.638;P=0.000)结果(除CD4+/CD8+比值外)的P值均小于0.001(P<0.001),有统计学意义。说明自然沉降法细胞自然形态保存良好,在高倍镜下识别细胞无误,准确度高。另外,细胞洗涤、抗原-抗体结合反应离心速度对获得显微镜下淋巴细胞红细胞花环形态的最佳效果有较大影响。结论:实用Mc Ab-A-E直接法检验淋巴细胞亚群时,自然沉降法分离PBMC效果优于淋巴细胞分离液梯度密度离心法。

Objective： Study the different methods of preparing cell suspensions and ceil natural form in practical McAb-A-E direct method. Methods： Select 20 cases of clinical specimens, prepare mononuclear cell suspension using natural sedimentation method （without any substance） or lymphocyte separation medium density gradient centrifugation, then using practical McAb-A-E direct method test and compare CD3^＋,CD4^＋,CD8^＋,CD4^＋/CD8^＋ and non-lymphocyte. Results： The experimental results analyzed by using paired test in SPSS13.0 statistical software.between the two groups CD3^＋（t=-4.197;P=0.000）,CD4^＋（t=-5.177;P=0.000）,CD8^＋（t=-4.24; P=0.000）,CD4^＋/CD8^＋（t=-1.758;P=0.095）and non-lymphocyte （t=-5.638;P=0.000）Results （besides CD4^＋/CD8^＋） all had statistical significance （P〈0.01）. To explain natural sedimentation method to preserve well cell natural form,in high power lens can correct recognition cell.In addition, washed cells, antigen antibody reaction after the centrifuge speed and time also had a great effect on the morphology of the lymphocytes. Conclusions： practical McAb-A-E direct method testing T-lymphocyte subsets used that natural sedimentation method （without any substance）to separate PBMC result that lymphocyte separation medium density gradient centrifugation.