用DNA-PCR方法于基因组水平检测K562细胞及慢性粒细胞白血病患者bcr/abl融合基因

Detection of bcr/abl fusion gene from K562 cell line and mononuclear cells of CML patients by DNA-PCR

目的:在慢性粒细胞白血病(慢粒)细胞系K 562及慢粒患者中尝试基于DNA水平的bcr/ab l融合基因检测方法。方法:以W aller的“L-T DNA PCR”方法为基础,并针对其引物位置设计的缺陷,改进设计了一套DNA-PCR引物,分别提取K 562细胞及慢粒患者外周血单个核细胞DNA,进行bcr/ab l融合基因扩增,并对扩增片段进行序列测定。结果:运用DNA-PCR方法成功地扩增出了K 562细胞及2名CM L患者外周血基因组的bcr/ab l融合基因片段,随后进行的片段测序进一步明确了融合基因的断裂位点。结论:DNA-PCR提供了一种基于基因组DNA水平的新检测手段,结合测序能鉴定融合基因的断裂位点,若结合定量PCR的方法则能监测患者的治疗效果与预后,还能检测患者经化疗后的残留微小病灶。

Objective： To detect bcr/abl fusion gene at DNA level in K562 cell line and mononuclear cells from patients with chronic myelogenous leukemia （CML）. Methods： Based on previous research, a set of DNA-PCR primers was redesigned. DNA from K562 cells and mononuclear cells of CML patients was extracted respectively. After DNA-PCR for bcr/abl fusion gene the amplified fragments were then sequenced. Results： At DNA level the bcr/abl fusion gene in K562 cells and mononuclear cells of 2 CML patients was amplified. Furthermore, the DNA brea fragments was fusion without kpoint of fusion gene in the above samples through sequencing of amplified localized. Conclusion： DNA-PCR offers a new detection technology for bcr/abl the expression of fusion gene.