CAJ | 학술논문

目的探讨半巢式甲基化特异性PCR( hn-MSP)的特性并了解p15基因甲基化和缺失在急性淋巴细胞白血病(ALL)发病中可能起到的作用.方法运用hn-MSP方法和基因组硫化修饰PCR( BSP)后直接测序方法,分析在25例初诊或复发不同阶段ALL患者以及部分恶性血液病细胞株中p15基因的甲基化和缺失状态,以10名健康者或非恶性血液病患者为对照组.以Molt-4细胞株为阳性对照,以对照组单个核细胞为阴性对照,分析hn-MSP方法的敏感性和特异性.结果 hn-MSP产物克隆测序结果与BSP结果一致,hn-MSP检测p15基因甲基化敏感性可达到1×10-5.25例ALL患者p15基因甲基化发生率为68.0％(17例)；25例ALL患者中3例p15基因外显子1缺失.对照组p15基因无甲基化或缺失.结论 p15基因甲基化状态在ALL患者中有较高的检出率,hn-MSP对分析p15基因甲基化状态具有较高的特异性和敏感性.
목적 탐토반소식갑기화특이성PCR( hn-MSP)적특성병료해p15기인갑기화화결실재급성림파세포백혈병(ALL)발병중가능기도적작용.방법 운용hn-MSP방법화기인조류화수식PCR( BSP)후직접측서방법,분석재25례초진혹복발불동계단ALL환자이급부분악성혈액병세포주중p15기인적갑기화화결실상태,이10명건강자혹비악성혈액병환자위대조조.이Molt-4세포주위양성대조,이대조조단개핵세포위음성대조,분석hn-MSP방법적민감성화특이성.결과 hn-MSP산물극륭측서결과여BSP결과일치,hn-MSP검측p15기인갑기화민감성가체도1×10-5.25례ALL환자p15기인갑기화발생솔위68.0％(17례)；25례ALL환자중3례p15기인외현자1결실.대조조p15기인무갑기화혹결실.결론 p15기인갑기화상태재ALL환자중유교고적검출솔,hn-MSP대분석p15기인갑기화상태구유교고적특이성화민감성.
Objective To explore the characteristics of hemi-nested methylation specific polymerase chain reaction (hn-MSP) and to find out the possible relationship between patterns of methylation or deletion and the developmet of adult acute lymphoblastic leukemia(ALL).Methods hn-MSP and bisulfit-sequencing PCR (BSP) were designed and adopted to analyze p15 gene methylation or deletion patterns in 25 adult ALL patients,malignant hematopathy cell lines and normal lymphocytes. hn-MSP and BSP products were cloned and sequenced.The sensitivity and specificity of hn-MSP were also analized.Results The sequencing results of hn-MSP and BSP products were consistent, and the sensitivity of detection of p15 methylation was up to 1.0×10-5.17 adult ALL patients (68 ％) were p15 gene hypermethylation and 3 patients were with deletion of p15 gene exon 1.There were no hypermethylation or deletion in the 10 controls.Conclusions The detection rate of p15 methylation in many tumors,especially in adult ALL,is frequent high.hn-MSP is highly sensitive and specific in analyzing p15 methylation.