流式免疫微球法快速、简易检测PML-RARα融合蛋白

niwanmao

通过流式免疫微球法快速、简易检测PML-RARα融合蛋白诊断急性早幼粒细胞白血病

这是leukemia杂志今年新出的文章，急性早幼粒细胞白血病（APL）也就是M3，其中97％的患者具有PML-RARα融合蛋白，该疾病如不及时治疗，会出现危及生命的出血。本文中作者采用流式免疫微球包裹法，可快速（约4－5小时）而敏感（可检测正常背景下10％的白血病细胞）地检测细胞裂解物中的PML-RARα融合蛋白。在163例初诊患者中（包括46例APL）中，与PML-RARα的PCR结果一致，而且检测结果与PML基因的断裂点位置无关。
在此，我想到，那些开发试剂的大公司，如BD、COULTER、DAKO等是不是能够及时开发出这种抗体，为临床造福。因为M3患者非常容易发生DIC，用流式微球法检测时间比PCR更快，而越早诊断M3，对临床治疗M3和患者的预后越有帮助。

Flow cytometric immunobead assay for fast and easy detection of PML–RARA fusion proteins for the diagnosis of acute promyelocytic leukemia
Leukemia (2012) 26, 1976–1985;
The PML–RARA fusion protein is found in approximately 97% of patients with acute promyelocytic leukemia (APL). APL can be associated with life-threatening bleeding complications when undiagnosed and not treated expeditiously. The PML–RARA fusion protein arrests maturation of myeloid cells at the promyelocytic stage, leading to the accumulation of neoplastic promyelocytes. Complete remission can be obtained by treatment with all-trans-retinoic acid (ATRA) in combination with chemotherapy. Diagnosis of APL is based on the detection of t(15;17) by karyotyping, fluorescence in situ hybridization or PCR. These techniques are laborious and demand specialized laboratories. We developed a fast (performed within 4–5 h) and sensitive (detection of at least 10% malignant cells in normal background) flow cytometric immunobead assay for the detection of PML–RARA fusion proteins in cell lysates using a bead-bound anti-RARA capture antibody and a phycoerythrin-conjugated anti-PML detection antibody. Testing of 163 newly diagnosed patients (including 46 APL cases) with the PML–RARA immunobead assay showed full concordance with the PML–RARA PCR results. As the applied antibodies recognize outer domains of the fusion protein, the assay appeared to work independently of the PML gene break point region. Importantly, the assay can be used in parallel with routine immunophenotyping for fast and easy diagnosis of APL.

M3患者的融合基因及其变异分布：

M3患者的融合基因及其变异分布

检测原理：

检测原理

初诊AML患者的检测结果（可见本方法对APL非常有特异性）：

初诊AML患者的检测结果

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