HpaⅡ—PCR检测急性白血病降钙素基因高度甲基化

Use of the HpaⅡ-PCR to Detect Hypermethylation of the Calcitonin Gene in Acute Leukemia

应用聚合酶链反应(PCR)结合HpaⅡ限制性内切酶消化法(HpaⅡ—PCR)检测78例急性白血病(AL)患者降钙素(CT)基因的甲基化快态。病例组待检细胞DNA经 HpaⅡ消化后,再用两对CT基因特异性引物作 PCR扩增,分别产生长度为 566 bp和1.4 kb特异片段。急性淋巴细胞白血病阳性率 68.8％(33/48),急性非淋巴细胞白血病73.3％(22/30),敏感性达10^(-3)。证明CT基因5’高度甲基化是白血病细胞克隆的特异标志。本方法对恶性血液病的诊断治疗及其微小残留病(MRD)的检测都具有重要意义。

The patterns of hypermethylation in calcitonin (CT) gene of 78 cases with acuteleukemia were detected by using polymerase chain reaction (PCR) in combination withHpa Ⅱ restriction endonuclease digestion. The DNA of the cells in the case ?oup to be de-tected tvere digested hy using Hpa Ⅱ, which then were amplified by using ?wo pairs ofprimers specific for the CT genes.The fragments of 566 bp and 1.4 kb were developed re-spectively. The positive rate of acute lymphoblastic leukemia and acute non -- lymphoblasticleukemia was 68. 8% (33/48) and 73. 3% (22/30) respectively the sensitivity was 3logs. This study suggested that hypermethylation of the calci?onin gene 5' -- flanking was thespecific molecular marker for the leukemic cell colony, and this approach may be clinically useful for the detection of malignant hematologic disorders and minimal residual disease(MRD).