HpaⅡ-PCR检测小儿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５８例急性白血病（ＡＬ）患儿降钙素（ＣＴ）基因的甲基化状态。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５６６ｂｐ和１．４ｋｂ特异片段。急性淋巴细胞白血病阳性率７１．４％（２５／３５），急性非淋巴细胞白血病７８．２％（１８／２３）。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。本课题受卫生部。

The patterns of hypermethylation in calcitonin(CT)gene of 58 cases with acute leukemia were detected by using polymerase chain reaction (PCR) in combination with HpaⅡ restriction endonuclease digestion .The DNA of the cells in the case group to be detected were digested by using HpaⅡ,which then were amplified by using two pairs of primers specific for the CT genes.The fragments of 566 bp and 1 4 kb were produced respectively.The positive rate of acute lymphoblastic leukemia was 71 4(25/35),and acute non lymphoblastic leukemia 78 2(18/23),the sensitivity being 3 logs.This study suggests that hypermethylation of the calcitonin gene 5′ flanking is the specific molcular marker for the leukemic cell colony.