DNA片段长度分析检测急性髓性白血病CEBPA基因突变

DNA fragment length analysis for screening of CEBPA mutations in acute myeloid leukaemia

目的建立检测急性髓性白血病(AML)髓系转录因子CCAAT增强子结合蛋白-A(CEBPA)基因突变的快速筛查方法。方法采用聚合酶链式反应(PCR)扩增产物片段长度分析及序列分析方法检测107例初治AML患者CEBPA基因全部编码区突变情况。来自同种基因移植供者的38例正常人作为对照。结果同时用测序及片段长度分析的方法检测了107例AML患者及38例正常人CEBPA突变的情况,在排除了已知的多态位点以后,在107例AML患者中筛查出22例患者存在CEBPA突变,其中4例患者存在2种CEBPA突变,18例患者存在1种CEBPA突变,而在38例正常人中没有发现CEBPA突变。与测序结果比较,片段长度分析检测CEBPA突变是敏感和有效的。结论片段长度分析是一种快速、敏感地筛查CEBPA基因突变的方法,适合常规的AML诊断。

Objective To establish a rapid screening method for CEBPA（CCAAT-Enhancer Binding Protein-alpha,CEBPA）mutations.Methods We detected mutations throughout the entire coding region of the CEBPA gene in 107 untreated AML patients using polymerase chain reaction （PCR） followed by sequence analysis and fragment length analysis.Thirty-eight normal donors from the allogeneic stem cell transplantation were served as control.Results We evaluated the approach by analysing 107 AML patient and 38 normal samples by both fragment length analysis and nucleotide sequencing.Twenty-two cases with CEBPA mutation were found in 107 AML patients after excluding all known CEBPA polymorphisms.Of 4 CEBPA mutated patients had two mutations and 18 patients had a single mutation.No mutation was found in 38 normal samples.CEBPA mutation detection by fragment length analysis is sensitive and efficient when compared to nucleotide sequencing.Conclusions We established a fast and sensitive CEBPA mutation screening method for routine AML diagnostics.