摘要
目的应用变性高效液相色谱(DHPLC)结合甲基化特异性PCR(MSP),建立一种临床可行的MGMT基因启动子甲基化检测方法,并探讨其灵敏度和特异性。方法选取MGMT甲基化标准品、30例脑胶质瘤组织和5例正常脑组织作为研究对象,分别应用MSP和MSP-DHPLC方法进行MGMT基因启动子区甲基化检测,比较两种方法的检测灵敏度和肿瘤组织甲基化检测率。结果标准品MGMT甲基化检测,MSP-DHPLC检测灵敏度为1%;MSP检测灵敏度为10%。30例胶质瘤组织用常规MSP和MSP-DHPLC检测,发生甲基化的样本例数分别为15(50%)和24(80%),其中9例样本MSP检测为非甲基化,而MSP-DHPLC检测为甲基化。5例正常脑组织采用两种方法检测均为非甲基化。结论 MSP-DHPLC方法检测MGMT基因甲基化灵敏度高于常规MSP。
Objective To develop a clinical feasible method to detect promoter methylation of MGMT by MSP-DHPLC and assess its sensitivity and specificity. Methods Methylation status of MGMT gene were detected using MSP and MSP-DHPLC methods respectively in 30 glioma tumor tissues, 5 non-tumor brain tissues, and 2 standard DNA with the purpose to compare the sensitivity of two detecting methods. Results The sensitivity of detecting MGMT gene methylation with MSP and MSP-DHPLC was 10% and 1% respectively. MGMT gene methylation was observed in 15（50%） tumor tissues with MSP and in 24（80%） tumor tissues with MSP-DHPLC, among which there are 9 samples can only detected through MSP-DHPLC but not MSP method. MGMT gene unmethylation were found in all non-tumor brain tissues detecting by both methods. Conclusion Detecting MGMT methylaiton with MSP-DHPLC method was more sensitive than MSP, indicating MSP-DHPLC may be a useful tool in clinical test ofMGMT gene rnethylation.
出处
《分子诊断与治疗杂志》
2011年第1期6-9,共4页
Journal of Molecular Diagnostics and Therapy
