Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Blood. 2012 Oct 11;120(15):e54-9. doi: 10.1182/blood-2012-05-434266. Epub 2012 Aug 29.
A 2-day method using flow cytometry and FISH for interphase cells was developed to detect monosomy 7 cells in myelodysplastic syndrome patients. The method, Interphase Chromosome Flow-FISH (IC Flow-FISH), involves fixation of leukocytes from blood, membrane permeabilization, hybridization of cellular DNA with peptide nucleic acid probes with cells intact, and analysis by flow cytometry. Hundreds to thousands of monosomy 7 cells were consistently detected from 10-20 mL of blood in patients with monosomy 7. Proportions of monosomy 7 cells detected in IC Flow-FISH were compared with results from conventional cytogenetics; identification of monosomy 7 populations was verified with FACS; and patient and donor cells were mixed to test for sensitivity. IC Flow-FISH allows for detecting monosomy 7 without requiring bone marrow procurement or the necessity of metaphase spreads, and wider applications to other chromosomal abnormalities are in development.
我们开发了一种使用流式细胞术和荧光原位杂交(FISH)技术检测骨髓增生异常综合征患者间期细胞单体 7 号染色体的 2 天方法。该方法称为间期染色体流式-FISH(IC Flow-FISH),包括从血液中固定白细胞、细胞膜通透化、细胞完整状态下用肽核酸探针与细胞 DNA 杂交,以及通过流式细胞术进行分析。在单体 7 号染色体患者的 10-20ml 血液中,我们可以从数百到数千个间期细胞中检测到单体 7 号染色体。IC Flow-FISH 检测到的单体 7 号染色体细胞的比例与常规细胞遗传学的结果进行了比较;用 FACS 验证了单体 7 号染色体细胞群体的鉴定;并混合患者和供体细胞以测试其灵敏度。IC Flow-FISH 无需采集骨髓或制备中期分裂象,即可检测单体 7 号染色体,并且正在开发更广泛地应用于其他染色体异常的方法。