BackgroundDetection of del(17p) in myeloma is generally performed by fluorescence in situ hybridization (FISH) on a slide with analysis of up to 200 nuclei. The small cell sample analyzed makes this a low precision test. We report the utility of an automated FISH method, called “immuno‐flowFISH”, to detect plasma cells with adverse prognostic risk del(17p) in bone marrow and blood samples of patients with myeloma.MethodsBone marrow (n = 31) and blood (n = 19) samples from 35 patients with myeloma were analyzed using immuno‐flowFISH. Plasma cells were identified by CD38/CD138‐immunophenotypic gating and assessed for the 17p locus and centromere of chromosome 17. Cells were acquired on an AMNIS ImageStreamX MkII imaging flow cytometer using INSPIRE software.ResultsChromosome 17 abnormalities were identified in CD38/CD138‐positive cells in bone marrow (6/31) and blood (4/19) samples when the percent plasma cell burden ranged from 0.03% to 100% of cells. Abnormalities could be identified in 14.5%–100% of plasma cells.ConclusionsThe “immuno‐flowFISH” imaging flow cytometric method could detect del(17p) in plasma cells in both bone marrow and blood samples of myeloma patients. This method was also able to detect gains and losses of chromosome 17, which are also of prognostic significance. The lowest levels of 0.009% (bone marrow) and 0.001% (blood) for chromosome 17 abnormalities was below the detection limit of current FISH method. This method offers potential as a new means of identifying these prognostically important chromosomal defects, even when only rare cells are present and for serial disease monitoring.

中文翻译：

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成像流式细胞术检测多发性骨髓瘤骨髓及循环浆细胞中del(17p)

背景骨髓瘤中 del(17p) 的检测通常通过载玻片上的荧光原位杂交 (FISH) 进行，并分析多达 200 个细胞核。分析的小细胞样本使得该测试的精度较低。我们报告了一种称为“免疫流 FISH”的自动化 FISH 方法的实用性，用于检测骨髓瘤患者的骨髓和血液样本中具有不良预后风险 del(17p) 的浆细胞。n= 31) 和血液 (n= 19) 使用免疫流 FISH 分析了 35 名骨髓瘤患者的样本。通过 CD38/CD138 免疫表型门控鉴定浆细胞，并评估 17 号染色体的 17p 位点和着丝粒。使用 INSPIRE 软件在 AMNIS ImageStreamX MkII 成像流式细胞仪上采集细胞。结果在 CD38/CD138 阳性细胞中鉴定出染色体 17 异常在骨髓 (6/31) 和血液 (4/19) 样本中，浆细胞负荷百分比范围为细胞的 0.03% 至 100%。14.5%~100%的浆细胞可检出异常。结论“免疫流FISH”成像流式细胞术可以检测骨髓瘤患者骨髓和血液样本中浆细胞的del(17p)。该方法还能够检测 17 号染色体的增加和丢失，这也具有预后意义。17号染色​​体异常的最低水平为0.009%（骨髓）和0.001%（血液），低于当前FISH方法的检测限。该方法具有作为识别这些对预后重要的染色体缺陷的新方法的潜力，即使仅存在稀有细胞并用于连续疾病监测。