摘要
目的:建立甲基化特异性PCR(MSP)技术检测死亡相关蛋白激酶(DAPK)基因启动子甲基化。方法:设计针对DAPK基因启动子的MSP引物,建立MSP检测体系对甲基化阳性模板进行检测,评价其特异性、重复性和灵敏性,并经测序鉴定。结果:所建立的甲基化MSP方法仅能扩增出甲基化阳性模板,不能扩增出未甲基化阳性模板和未硫化处理的DNA模板;对不同梯度稀释的甲基化阳性模板进行检测,其最大敏感性可达2%;在不同时间进行甲基化MSP检测的结果均一致。结论:成功建立的DAPK基因启动子甲基化MSP检测方法具有良好的特异性、灵敏性和重复性,可用于肿瘤标本的批量检测。
Objective: To establish the methylation-specific PCR(MSP) for the detection of promoter methylation of death-associated protein kinase(DAPK) gene.Methods: The primers of MSP aiming at the DAPK promoter were designed.MSP was established to detect the methylation positive DNA templates.The specificity,sensibility and repeatability were evaluated. Results: The established methylated MSP could only amplify the methylation positive DNA template,but not the unmethylated DNA template and unmodified DNA template.In a series of dilution,methylated MSP could detect 2% methylation positive DNA templates.The amplified results of same sample were identical on different time. Conclusion: The methylated MSP technique for detecting of DAPK methylation was established successfully and can be used for analyzing samples from cancer patients.
出处
《江苏大学学报(医学版)》
CAS
2009年第1期23-26,共4页
Journal of Jiangsu University:Medicine Edition
基金
江苏省医学重点人才资助项目(RC2007035)
江苏大学临床医学科技发展基金资助项目(JLY20080048)
镇江市社会发展基金资助项目(SH2006032)