根据更新的指南，基于 FLAER 的检测提高了 PNH 克隆检测的灵敏度,Indian Journal of Hematology and Blood Transfusion

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根据更新的指南，基于 FLAER 的检测提高了 PNH 克隆检测的灵敏度
Indian Journal of Hematology and Blood Transfusion ( IF 0.7 ) Pub Date : 2019-11-05 , DOI: 10.1007/s12288-019-01220-8
Prabhu Manivannan ₁ , Seema Tyagi ₂ , Hara Prasad Pati ₂ , Renu Saxena ₂

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流式细胞术已成为检测阵发性夜间血红蛋白尿 (PNH)、再生障碍性贫血 (AA) 和骨髓增生异常综合征 (MDS) 异常克隆的“金标准”。这项试点研究于 2015 年进行，主要目的是评估基于单管荧光气溶素 (FLAER) 的测试的效用及其与两种非基于 FLAER 的测试（CD55、CD59、CD24 和 CD66b）在检测异常 PNH 克隆方面的比较在这些新诊断的病例中。次要目的是尝试根据诊断时的临床、实验室特征和克隆大小将 PNH 与与 PNH 克隆相关的 AA/MDS 病例区分开来。在本研究中，在所有 PNH 病例（n = 12）中，使用基于单管 FLAER 的测试和两管非基于 FLAER 的测试检测到异常 PNH 克隆，健康受试者 (n = 18) 和具有 PNH 克隆的 AA/MDS (n = 9)，并与诊断时的临床和实验室特征进行比较。接受者操作曲线 (ROC) 分析定义了粒细胞 (> 0.7%) 和单核细胞 (> 0.9%) 中 FLAER 的最佳截止值。在这些细胞中，基于 FLAER 和非 FLAER 的测试之间存在显着的正相关（r > 0.3 和 p < 0.05）。基于 FLAER 的测试帮助我们在 4 名患者中找到了后一种技术遗漏的较小克隆，从而提高了其敏感性，并且在技术上也证明是具有成本效益的（1800 卢比对 2100 卢比）。即使在 PNH 患者中，与非基于 FLAER 的抗体组相比，使用 FLAER 的克隆大小也略高。在PNH和AA/MDS组中，单核细胞的克隆大小始终高于粒细胞。诊断时的骨髓细胞构成和粒细胞和单核细胞克隆的平均大小在这些组之间显示出显着的统计学显着'p'值<0.0001。在这项初步研究中，基于单管 FLAER 的 PNH 测试改进了所有 PNH、AA/MDS 和 PNH 克隆病例的克隆检测。大多数 PNH 病例的克隆大小 > 30%，而在 AA/MDS 中，诊断时通常 < 10%。因此，这种较新的技术不仅提高了 PNH 克隆检测的灵敏度，而且被证明是具有成本效益的。在大多数 PNH 病例中为 30%，而在 AA/MDS 中，诊断时通常<10%。因此，这种较新的技术不仅提高了 PNH 克隆检测的灵敏度，而且被证明是具有成本效益的。在大多数 PNH 病例中为 30%，而在 AA/MDS 中，诊断时通常<10%。因此，这种较新的技术不仅提高了 PNH 克隆检测的灵敏度，而且被证明是具有成本效益的。

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FLAER Based Assay According to Newer Guidelines Increases Sensitivity of PNH Clone Detection

Flow cytometry has become ‘gold standard’ for detecting abnormal clones in paroxysmal nocturnal hemoglobinuria (PNH), aplastic anemia (AA) and myelodysplastic syndrome (MDS). This pilot study was conducted in 2015 with a primary aim to evaluate the utility of single tube fluorescent aerolysin (FLAER) based testing and its comparison with two tubes non-FLAER based testing (CD55, CD59, CD24 and CD66b) in detecting abnormal PNH clones in these newly diagnosed cases. The secondary aim was an attempt to distinguish PNH from AA/MDS cases associated with PNH clones based on clinical, laboratory features and clone size at diagnosis. In this study, the abnormal PNH clones were detected using a single tube FLAER based testing and two tubes non-FLAER based testing in all cases of PNH (n = 12), healthy subjects (n = 18) and AA/MDS with PNH clone (n = 9) and compared with clinical and laboratory features at diagnosis. The receiver operator curve (ROC) analysis defined the optimal cut-offs for FLAER in granulocytes (> 0.7%) and monocytes (> 0.9%). There was significant positive correlation between FLAER and non-FLAER based testing in these cells (r > 0.3 and p < 0.05). FLAER based testing helped us in picking up smaller clones which were missed by latter technique in four patients thereby increasing its sensitivity and also technically proved to be cost-effective (Rs. 1800 vs. Rs. 2100). Even in PNH patients, the clone size was slightly higher by using FLAER when compared to non-FLAER based antibodies panel. The clone size of monocytes was always higher than granulocytes in both PNH and AA/MDS groups. Bone marrow cellularity and mean size of granulocytes and monocytes clone at diagnosis showed a striking statistically significant ‘ p ’ value of < 0.0001 between these groups. In this pilot study, a single tube FLAER based PNH testing had improved clone detection in all cases of PNH, AA/MDS with PNH clones. The clone size was > 30% in majority of PNH cases whereas in AA/MDS, it was usually < 10% at diagnosis. Hence this newer technique not only increased the sensitivity of PNH clone detection but also proved to be cost-effective.

更新日期：2019-11-05

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