摘要
目的 通过对 98例献血者外周血HLA B2 7的检测 ,建立并优化测定HLA B2 7的流式细胞术 ,并评价B2 7在早期辅助临床诊断以强直性脊柱炎为代表的脊柱关节病中的应用价值。方法 分别采用EDTA和肝素钠两种不同的抗凝剂 ,以及加单抗不同的量 ( 7～ 3 0 μl)检测 2个B2 7阳性和 9个B2 7阴性个体的全血 ,采用EDTA抗凝和 15 μl的单抗剂量检测献血者外周血 ,探寻流式细胞术检测HLA B2 7的最佳实验条件 ;另以微量淋巴细胞毒实验分别对 2例阳性标本和 9例阴性标本进行验证。结果 通过微量淋巴细胞毒和流式细胞术两种方法检测 98例献血者外周血HLA B2 7中 2例为阳性 ,96例为阴性 ,且EDTA和肝素钠两种不同的抗凝剂 ,以及加单抗不同的量 ( 7～ 3 0 μl)对流式细胞术检测HLA B2 7无任何影响。结论 优化并建立了一种简便省时、稳定性和重复性皆好的流式细胞仪用于检测HLA B2 7的方法 ;HLA B2 7与强直性脊柱炎高度相关 ,HLA B2 7检测已成为临床支持以强直性脊柱炎为首的脊柱关节病的诊断与鉴别诊断的重要依据 ,具有重要的临床意义。
Objective To establish and optimize flow cytometry method for the detection of human leukocyte antigen B27 (HLA B27) and compare it with serological method. Methods A total of 98 samples from blood donors were entered into the study. HLA B27 was detected with ethylenediaminetetra aceticacid(EDTA), heparin sodium and different dosages of B27 monoclonal antibody, respectively. Other 30 samples were typed by microlymphocytotoxicity test(serological method). Results Genotyping for HLA B27 was successfully performed in 98 samples by flow cytometry, containing 2 cases with B27 positive, 96 cases with B27 negative. The types of anticoagulant agents and dosages of monoclonal antibody(7～30 μl) had no remarkable effect on the results. The results of serological method for the detection of 30 samples were consistent with those by flow cytometry method. Conclusion Genotyping for HLA B27 by flow cytometry is a rapid and accurate technique with high sensitivity, suitable for clinical application. Detection of HLA B27, a very important basis for the diagnosis and differential diagnosis of ankylosing spondylitis(AS) and other joint diseases of the vertebral column, is of a remarkable clinical significance.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2003年第8期727-729,共3页
Journal of Third Military Medical University
