Halifax-FCXM 流式细胞仪快速优化 96 孔板交叉配型与 Luminex 单抗原检测的评估
Assessment of Rapid Optimized 96-well Tray Flow Cytometric Crossmatch (Halifax-FCXM) with Luminex Single Antigen Test.
机构信息
Department of Laboratory Medicine, Incheon St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Republic of Korea.
Department of Laboratory Medicine, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Republic of Korea.
出版信息
Hum Immunol. 2021 Apr;82(4):302-308. doi: 10.1016/j.humimm.2021.02.003. Epub 2021 Mar 18.
INTRODUCTION
Flow cytometric crossmatch assay (FCXM) is a sensitive cell-based method for evaluating the presence of donor-specific antibodies (DSA) before transplantation. Recently, 96-well tray FCXM protocol (Halifax FCXM) with improved test efficiency has been introduced. The objective of the present study was to assess the performance of Halifax FCXM by correlating with DSA results based on single antigen bead (SAB) assays (virtual crossmatch, VXM).
METHODS
A total of 341 FCXMs were evaluated for the detection of HLA-DSA. A positive VXM was defined as having at least one HLA - DSA (HLA-A, B, Cw, DR, DQB1) with ≥ 1000 MFI (mean fluorescence intensity) identified by SAB assay.
RESULTS
Of a total 341 cases, 113 showed class I VXM (+) with class I DSA MFI ≥ 1000 exclusively against one or more donor HLA class I antigens (HLA-A, B, Cw), 72 had class I-/II + DSA, and 156 had VXM(-). Halifax T-FCXM showed a sensitivity of 87.6% (99/113) and a specificity of 98.2% (224/228) for detecting class I VXM (+). The concordance between T-FCXM and class I VXM was 94.7% (323/341). Halifax B-FCXM showed a sensitivity of 58.3% (42/72) and a specificity of 98.7% (154/156) for detecting class I-/II + DSAs. The concordance between B-FCXM and class I-/II + VXM was 86.0% (196/228). When we separately analyzed data, B-FCXM detected HLA-DR (+) (68.8%) and HLA-DQ (+) DSAs (71.0%) similarly (P > 0.05). T-FCXM detected 87.6%, 97.2%, and 98.2% of class I DSA-positive cases with MFI values (sumDSA) ≥ 1000, ≥ 3000, and ≥ 5000, respectively. B-FCXM detected 58.3% of class I-II + DSA -positive (≥1000) cases, but detected 76.7% (33/43) and 89.2% (33/37) of class I-II + DSAs if MFI values of sumDSA and immunodominant DSA (iDSA) were above 5000, respectively. Halifax FCXM had sensitivities of 91.5% and 96.2% for detecting VXM (+) having MFI values above 5000 for class I or class II sumDSA and iDSA, respectively.
CONCLUSION
Halifax FCXM showed a good correlation, especially with SAB assay-based high MFI DSA or sumDSA. Concurrent application of FCXM with VXM can improve pre-transplant risk assessment and progress organ allocation efficiency.
简介
流式细胞交叉配型检测（FCXM）是一种用于评估移植前供体特异性抗体（DSA）存在的敏感细胞检测方法。最近，一种改进了检测效率的 96 孔板 FCXM 方案（哈利法克斯 FCXM）被引入。本研究的目的是通过与基于单抗原珠（SAB）检测的虚拟交叉配型（VXM）的相关性来评估哈利法克斯 FCXM 的性能。
方法
共评估了 341 例 FCXM 以检测 HLA-DSA。将至少有一种 HLA-A、B、Cw、DR、DQB1 与≥1000 MFI（平均荧光强度）的 HLA-DSA（阳性 VXM）定义为 SAB 检测阳性。
结果
在总共 341 例中，113 例显示 I 类 VXM（+），其中 I 类 DSA MFI≥1000 仅针对一个或多个供体 HLA I 类抗原（HLA-A、B、Cw），72 例显示 I-/II 类 DSA（+），156 例显示 VXM（-）。哈利法克斯 T-FCXM 检测 I 类 VXM（+）的敏感性为 87.6%（99/113），特异性为 98.2%（224/228）。T-FCXM 与 I 类 VXM 的一致性为 94.7%（323/341）。哈利法克斯 B-FCXM 检测 I-/II 类 DSA（+）的敏感性为 58.3%（42/72），特异性为 98.7%（154/156）。B-FCXM 与 I-/II 类 DSA 的一致性为 86.0%（196/228）。当我们分别分析数据时，B-FCXM 检测 HLA-DR（+）（68.8%）和 HLA-DQ（+）DSA（71.0%）的敏感性相似（P>0.05）。T-FCXM 分别检测到具有 MFI 值（sumDSA）≥1000、≥3000 和≥5000 的 I 类 DSA 阳性病例的 87.6%、97.2%和 98.2%。B-FCXM 检测到 58.3%的 I 类-II 类 DSA（+）阳性（≥1000）病例，但如果 sumDSA 和免疫显性 DSA（iDSA）的 MFI 值高于 5000，则分别检测到 76.7%（33/43）和 89.2%（33/37）的 I 类-II 类 DSA。哈利法克斯 FCXM 对 MFI 值大于 5000 的 I 类或 II 类 sumDSA 和 iDSA 的 VXM（+）检测的敏感性分别为 91.5%和 96.2%。
结论
哈利法克斯 FCXM 与基于 SAB 检测的高 MFI DSA 或 sumDSA 相关性良好。FCXM 与 VXM 的同时应用可以提高移植前的风险评估和器官分配效率。
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