Reynolds W M, Evans P R, Lane A C, Howell W M, Wilson P J, Wong R, Smith J L
Wessex Histocompatibility Group, Southampton University Hospitals NHS Trust, Shirley, United Kingdom.
Cytometry. 1994 Jun 15;18(2):109-15. doi: 10.1002/cyto.990180210.
Tissue typing can help in the diagnosis of the seronegative arthropathy ankylosing spondylitis. Using an automatic sample preparation system and flow cytometry (FACSPrep/FACScan) we have developed a test for HLA-B27 screening using whole blood which is both rapid, reproducible, and permits simultaneous screening of large numbers of samples. We have used both indirect (248 cases) and direct (126 cases) monoclonal antibody staining techniques. Results were assessed using median channel shift (CS) from the negative control and relative fluorescence intensity. There is known cross-reactivity between HLA-B27 and HLA-B7 with the monoclonal antibody (MoAb) HLA-ABC-m3. Using this antibody and indirect staining, HLA-B7 samples had a significantly lower CS value than HLA-B27 samples. In 60% of these cases there was a clear distinction between HLA-B27 and HLA-B7. All HLA-B27 and HLA-B7 negative samples had a CS of 0 (P < 0.001). Using direct dual staining with CD3 and HLA-B27, all HLA-B27 and HLA-B7 negative samples had a low CS value (15 maximum), HLA-B7 samples had an intermediate CS value (20-85), and HLA-B27 samples had the highest CS values (70 upward). This system permits large scale rapid negative screening of samples with the elimination of over 60% as negatives (CS < 15). Limitations as a definitive test for HLA-B27 are due to MoAb cross-reactivity with HLA-B7 which necessitates other confirmatory techniques. The substitution of the HLA-ABC-m3 with the recently available HLA-B27 specific MoAb FD705 would substantially increase the value of this technique for routine HLA-B27 typing.
组织分型有助于血清阴性关节病强直性脊柱炎的诊断。我们使用自动样本制备系统和流式细胞仪（FACSPrep/FACScan）开发了一种全血HLA - B27筛查试验，该试验快速、可重复，且能同时筛查大量样本。我们使用了间接（248例）和直接（126例）单克隆抗体染色技术。通过与阴性对照的中位通道偏移（CS）和相对荧光强度来评估结果。已知HLA - B27与HLA - B7在单克隆抗体（MoAb）HLA - ABC - m3上存在交叉反应。使用该抗体和间接染色时，HLA - B7样本的CS值明显低于HLA - B27样本。在60%的此类病例中，HLA - B27和HLA - B7之间有明显区别。所有HLA - B27和HLA - B7阴性样本的CS值为0（P < 0.001）。使用CD3和HLA - B27的直接双重染色时，所有HLA - B27和HLA - B7阴性样本的CS值较低（最大为15），HLA - B7样本的CS值中等（20 - 85），而HLA - B27样本的CS值最高（70及以上）。该系统允许对样本进行大规模快速阴性筛查，可排除超过60%的阴性样本（CS < 15）。作为HLA - B27的确诊试验的局限性在于MoAb与HLA - B7的交叉反应，这需要其他确证技术。用最近可用的HLA - B27特异性MoAb FD705替代HLA - ABC - m3将大大提高该技术在常规HLA - B27分型中的价值。
