Levering Wilfried H B M, Sintnicolaas Kees, Wind Henk, Hooijkaas Herbert, Gratama Jan W
Laboratory for Histocompatibility and Immunogenetics, Sanquin Blood, Bank South West Region, Rotterdam, The Netherlands.
Curr Protoc Cytom. 2005 Aug;Chapter 6:Unit6.22. doi: 10.1002/0471142956.cy0622s33.
HLA-B27 is common to the entire group of seronegative spondyloarthropathies, and assessment of HLA-B27 is of diagnostic importance if any of these diseases are considered. Among the alternatives to traditional typing by the complement-dependent cytotoxicity assay, flow cytometric HLA-B27 screening is most widely used in general diagnostic laboratories, as it is simple, rapid, and cost effective. This unit describes screening for HLA-B27 on peripheral blood lymphocytes using more than one HLA-B27 monoclonal antibody to detect possible cross-reactivity with non-HLA-B27 antigens. Screening is hampered by the lack of true monospecific anti-HLA-B27 monoclonal antibodies. Cross-reactivities of anti-HLA-B27 with other HLA-B antigens have been reported. The authors recommend use of GS145.2 and FD705 antibodies, as this combination avoids most false-positive conclusions. Lyse-and-wash sample processing is employed. A multiparameter flow methodology is applied. Three to four parameters-forward scatter, side scatter, HLA-B27 expression, and, in some cases CD3 or B7 expression-are acquired.
HLA - B27在整个血清阴性脊柱关节病组中很常见，如果考虑这些疾病中的任何一种，HLA - B27的评估具有诊断重要性。在传统的补体依赖细胞毒性试验分型的替代方法中，流式细胞术HLA - B27筛查在一般诊断实验室中应用最为广泛，因为它简单、快速且具有成本效益。本单元描述了使用多种HLA - B27单克隆抗体对外周血淋巴细胞进行HLA - B27筛查，以检测与非HLA - B27抗原可能的交叉反应性。由于缺乏真正的单特异性抗HLA - B27单克隆抗体，筛查受到阻碍。有报道称抗HLA - B27与其他HLA - B抗原存在交叉反应性。作者建议使用GS145.2和FD705抗体，因为这种组合可避免大多数假阳性结果。采用裂解-洗涤样本处理方法。应用多参数流式方法。获取三到四个参数——前向散射、侧向散射、HLA - B27表达，以及在某些情况下的CD3或B7表达。
