Bonnaud G, Aupetit C, Preux P M, Cogné M, Drouet M
Laboratoire d'Immunologie et Immunogénétique, CHRU Dupuytren, Limoges, France.
Clin Rheumatol. 1999;18(1):23-7. doi: 10.1007/s100670050046.
HLA-B27 typing contributes to the diagnosis of ankylosing spondylitis. The classical technique of microlymphocytotoxicity is costly and can give false-negative results. We have compared 304 samples using two relatively new methods - flow cytometry and PCR-SSP - and evaluated their respective uses in routine analysis. Flow cytometric HLA-B27 testing was performed using three monoclonal anti-B27 antibodies (HLA-ABC-m3, GS145.2 and FD705 clones). Cut-off values were established to differentiate HLA-B27-positive from HLA-B27-negative samples with ROC curves. Although flow cytometric analysis with a reliable monoclonal antibody (mAb) is valuable for HLA screening, none of the HLA-B27 flow cytometry protocols was sufficient on its own to ascertain the HLA phenotype in 100% of samples. Two false negatives were observed with the FD705 mAb and the use of two different monoclonal antibodies did not increase the accuracy of HLA-B27 typing. HLA-B27 typing using molecular biology is a reliable but costly technique. Therefore we suggest that DNA typing could be used as a complementary technique and applied to samples whose HLA-B27 phenotype cannot be determined by flow cytometry. The association of flow cytometry and DNA typing is, in our experience, an economical and reliable approach.
HLA - B27分型有助于强直性脊柱炎的诊断。经典的微量淋巴细胞毒性技术成本高昂且可能产生假阴性结果。我们使用两种相对较新的方法——流式细胞术和聚合酶链反应 - 序列特异性引物法（PCR - SSP）对304个样本进行了比较，并评估了它们在常规分析中的各自用途。使用三种单克隆抗 - B27抗体（HLA - ABC - m3、GS145.2和FD705克隆）进行流式细胞术HLA - B27检测。通过ROC曲线确定临界值以区分HLA - B27阳性和HLA - B27阴性样本。尽管使用可靠的单克隆抗体（mAb）进行流式细胞术分析对于HLA筛查很有价值，但没有一种HLA - B27流式细胞术方案能够单独在100%的样本中确定HLA表型。使用FD705 mAb观察到两例假阴性，并且使用两种不同的单克隆抗体并未提高HLA - B27分型的准确性。使用分子生物学进行HLA - B27分型是一种可靠但成本高昂的技术。因此，我们建议DNA分型可作为一种补充技术，应用于那些无法通过流式细胞术确定HLA - B27表型的样本。根据我们的经验，流式细胞术和DNA分型相结合是一种经济且可靠的方法。
