急性髓系白血病17种基因异常的检测

Detection of 17 Kinds of gene abnormalities in acute myeloid leukemia

目的:探讨应用多重巢式RT-PCR、荧光定量PCR、PCR-SSCP银染技术检测初诊急性髓系白血病(acute myeloid leukemia,AML)中17种基因异常表达及在各亚型的分布情况,为个体化治疗提供依据。方法:采用多重巢式RT-PCR检测融合基因,PCR检测FLT3-ITD,荧光定量PCR检测NPM1的突变类型(A、B、D、I和R)及C-kit/D816V,PCR-SSCP银染技术检测CEBPA。对140例初诊AML患者(APL除外)的骨髓进行17种基因异常分析,并与骨髓染色体检测结果进行对比。结果:在140例初诊AML患者中检出基因异常占69例(49.3%),其中包括FLT3-ITD、NPM1、C-kit/D816V、CEBPA、HOX11、CBFβ/MYH11、AML-ETO、MLL/AF6、MLL/AF10、dup MLL、EVI1。同时对140例初治AML患者采用G显带技术行染色体核型分析,128例获得可供分析的染色体核型,其中57例(44.5%)检出染色体结构和数目异常。PCR在急性髓系白血病基因检测中较染色体核型分析具有更高的检出率。结论:PCR协同染色体核型分析检测初诊AML患者的基因异常,能提高临床诊断率、指导疾病危险度分组,为判断预后及监测微小残留病提供更好的理论依据。

Objective： To investigate the application of multiplex reverse transcription- polymerase chain reaction（ RT- PCR）,fluorescent quantitative polymerase chain reaction（ FQ- PCR）,polymerase chain reaction single stranded conformation polymorphism（ PCR- SSCP） and silver staining technique for the detection of 17 gene abnormalities in de novo acute myeloid leukemia（ AML）,and explore the expression and distribution among the subtypes of AML,and direct individual therapy. Methods： The bone marrow samples from 140 newly diagnosed AML patients（ APL except） were analyzed with multiplex nested RT- PCR to detect fusion genes,PCR to detect FLT3- ITD,FQ- PCR to detect NPM1（ A,B,D,I and R） and C- kit / D816 V,CEBPA was examined by PCR- SSCP and silver staining technique. In addition,chromosomal karyotypes were investigated. Results： Of the 140 leukemic samples,69（ 49. 3%） carried abnormal genes including FLT3- ITD,NPM1,C- kit / D816 V,CEBPA,HOX11,CBFβ / MYH11,AML- ETO,MLL / AF6,MLL / AF10,dup MLL,EVI1. In addition,chromosomal karyotypes by G banding techniques were examined in the 1 4 0 de novo AML,data were available in 1 2 8 patients and cytogenetic aberrations were detected in only 57（ 44. 5%） of them. The detection rate of PCR was higher than karyotype analysis in gene test of AML. Conclusion： Combined application of PCR and karyotype analysis for the detection of gene abnormalities in de novo AML,can improve the rate of clinical diagnosis,direct risk grouping,predict prognosis,and monitor minimal residual disease.