Detection of cytoplasmic nucleophosmin expression by imaging flow cytometry
Detection of cytoplasmic nucleophosmin expression by imaging flow cytometry
复制标题
成像流式细胞术检测细胞质核磷蛋白表达
作者：
L. Grimwade;E. Gudgin;D. Bloxham;G. Bottley;G. Vassiliou;B. Huntly;M. Scott;W. Erber
大约三分之一的急性髓系白血病 (AML) 病例会发生核磷蛋白 NPM1 基因突变。这些突变导致突变 NPM 蛋白在细胞质中积累。 NPM1突变目前通过分子方法检测。使用来自 37 名 AML 患者的样本，我们研究了成像流式细胞术是否可以成为当前技术的可行替代方案。骨髓/外周血细胞用抗 NPM 抗体和 DRAQ5 核染色剂进行染色，并在 ImageStream 成像流式细胞仪（Amnis Corp.，西雅图，美国）上获取数据。使用相似性特征进行数据分析，我们证明该技术可以基于细胞质 NPM 蛋白染色成功识别具有 NPM1 突变的 AML 病例（相似性阈值为 1.1，敏感性为 88%，特异性为 90%）。在 0-2 评分系统中结合平均荧光强度和不同染色百分比的数据进一步提高了灵敏度 (100%)。成像流式细胞术有可能成为标准流式细胞术抗体组的一部分，以识别潜在的 NPM1 突变，作为诊断和微小残留病监测的一部分。成像流式细胞术是一项令人兴奋的技术，在血液恶性肿瘤的诊断中具有许多可能的应用，包括整合模式的潜力。 © 2012 国际细胞计数促进会。
Mutations within the nucleophosmin NPM1 gene occur in approximately one‐third of cases of acute myeloid leukemia (AML). These mutations result in cytoplasmic accumulation of the mutant NPM protein. NPM1 mutations are currently detected by molecular methods. Using samples from 37 AML patients, we investigated whether imaging flow cytometry could be a viable alternative to this current technique. Bone marrow/peripheral blood cells were stained with anti‐NPM antibody and DRAQ5 nuclear stain, and data were acquired on an ImageStream imaging flow cytometer (Amnis Corp., Seattle, USA). Using the similarity feature for data analysis, we demonstrated that this technique could successfully identify cases of AML with a NPM1 mutation based on cytoplasmic NPM protein staining (at similarity threshold of 1.1 sensitivity 88% and specificity 90%). Combining data of mean fluorescence intensity and % dissimilar staining in a 0–2 scoring system further improved the sensitivity (100%). Imaging flow cytometry has the potential to be included as part of a standard flow cytometry antibody panel to identify potential NPM1 mutations as part of diagnosis and minimal residual disease monitoring. Imaging flow cytometry is an exciting technology that has many possible applications in the diagnosis of hematological malignancies, including the potential to integrate modalities. © 2012 International Society for Advancement of Cytometry.
影响因子：
2.9
作者：
DUMBAR, TS;GENTRY, GA;OLSON, MOJ
通讯作者：
OLSON, MOJ