Ward A M, Nikaein A
Immunology Department, Allied Clinical Laboratories, Hurst, Texas, USA.
Cytometry. 1995 Mar 15;22(1):65-9. doi: 10.1002/cyto.990220112.
HLA-B27 is an antigen associated with the disease ankylosing spondylitis. Ninety percent of Caucasians with ankylosing spondylitis possess the HLA-B27 antigen. However, only 20% of Caucasians with the HLA-B27 antigen will develop the disease. Defining the presence or absence of the HLA-B27 antigen can be helpful in differentiating ankylosing spondylitis from juvenile rheumatoid arthritis. In this study, we evaluated the application of two monoclonal antibodies (MoAbs), using flow cytometric analysis for the detection of HLA-B27 antigen. After an initial comparison of HLA-B27 analysis by flow cytometry to the standard microlymphocytotoxicity assay, cutoffs were established to differentiate HLA-B27 positive from HLA-B27 negative. One MoAb showed very reliable results with > 99% accuracy in discriminating HLA-B27 positive from HLA-B27 negative samples. Various parameters were investigated to obtain the optimum results and showed that incubation time, reagent lot, and the flow cytometric instrument can affect the results. We concluded that a reliable MoAb and flow cytometry are valuable for the rapid and inexpensive determination of HLA-B27 typing in the clinical setting. However, testing conditions can affect the accuracy of results; therefore, adequate parallel testing in various conditions must be performed in order to establish the proper standards.
HLA - B27是一种与强直性脊柱炎相关的抗原。90%的患有强直性脊柱炎的高加索人拥有HLA - B27抗原。然而，仅有20%拥有HLA - B27抗原的高加索人会患上这种疾病。确定HLA - B27抗原的存在与否有助于区分强直性脊柱炎和青少年类风湿性关节炎。在本研究中，我们评估了两种单克隆抗体（MoAbs）的应用，采用流式细胞术分析来检测HLA - B27抗原。在通过流式细胞术对HLA - B27进行分析并与标准微量淋巴细胞毒性试验进行初步比较后，确定了区分HLA - B27阳性和HLA - B27阴性的临界值。一种单克隆抗体显示出非常可靠的结果，在区分HLA - B27阳性和HLA - B27阴性样本方面准确率超过99%。为获得最佳结果，对各种参数进行了研究，结果表明孵育时间、试剂批次和流式细胞仪会影响结果。我们得出结论，一种可靠的单克隆抗体和流式细胞术对于在临床环境中快速且低成本地确定HLA - B27分型很有价值。然而，检测条件会影响结果的准确性；因此，必须在各种条件下进行充分的平行检测以建立适当的标准。
