The detection of HLA antibodies is important in clinical practice, such as platelet transfusion refractoriness and transfusion-related lung injury. However, difficulties are associated with the preparation of panel cells for conventional HLA detection systems using intact cells, such as the immunocomplex capture fluorescence analysis (ICFA).

Based on an ICFA analysis, HEK293 cells stably transfected with the HLA-A locus were used instead of peripheral blood mononuclear cells (PBMC). The reactivity, sensitivity, and stability of transfectants were examined.

All 20 antisera to HLA-A identified by LABScreen® Single Antigen class I (LS-SA1) were reactive to our modified-ICFA (m-ICFA) and showed the same specificities as those in LS-SA1, indicating the cell surface expression and correct antigenicity of the HLA-A locus in transfectants. The expression of HLA class I antigens was similar between transfectants frozen for 6 years and those prior to freezing. In the reaction of the anti-A24 or anti-A33 antibody vs each transfectant, the index of m-ICFA was higher than that of WAKFlow® ICFA. Our m-ICFA also showed that false negative reactions sometimes observed in capture assays may be avoided.

By using HLA-A transfectants as ICFA targets, we herein developed m-ICFA. Our m-ICFA may avoid false negative reactions of capture assay like enzyme-linked immunosorbent assay and can also be carried out in almost any laboratory without cell culture facilities.

HLA 抗体的检测在临床实践中很重要，例如血小板输注难治性和输血相关的肺损伤。然而，使用完整细胞（如免疫复合物捕获荧光分析 (ICFA)）为常规 HLA 检测系统制备面板细胞存在困难。

根据 ICFA 分析，使用 HLA-A 基因座稳定转染的 HEK293 细胞代替外周血单核细胞 (PBMC)。检查了转染子的反应性、敏感性和稳定性。

由 LABScreen® Single Antigen I 类 (LS-SA1) 鉴定的所有 20 种 HLA-A 抗血清都对我们的修饰的 ICFA (m-ICFA) 有反应，并显示出与 LS-SA1 相同的特异性，表明细胞表面表达和转染子中 HLA-A 基因座的正确抗原性。HLA I 类抗原的表达在冷冻 6 年的转染子和冷冻前的转染子之间相似。在抗 A24 或抗 A33 抗体与每种转染子的反应中，m-ICFA 的指数高于 WAKFlow® ICFA。我们的 m-ICFA 还表明可以避免有时在捕获分析中观察到的假阴性反应。

通过使用 HLA-A 转染子作为 ICFA 目标，我们在此开发了 m-ICFA。我们的 m-ICFA 可以避免酶联免疫吸附试验等捕获试验的假阴性反应，也可以在几乎任何没有细胞培养设施的实验室中进行。