甲基化特异性PCR的评价与优化  

The evaluation and optimization of methylation-specific polymerase chain reaction

作　　者：付祥胜[1] 张亚历[2] 李丽[1] 肖岚月[1] 邱野[1] 周贤[1] 张巍[1] 

机构地区：[1]四川省泸州医学院附属医院消化内科,646000 [2]南方医科大学南方医院消化疾病研究所,广州510515

出　　处：《检验医学与临床》2012年第15期1862-1863,1865,共3页Laboratory Medicine and Clinic

基　　金：泸州医学院自然科学基金资助课题(2010-108)

摘　　要：目的探讨甲基化特异性PCR(MSP)在DNA甲基化检测中的应用价值。方法以SHH基因启动子区全甲基化克隆和全非甲基化克隆(质粒)为模板,评价MSP的敏感性和特异性。结果 MSP能检测出样本中低含量的甲基化DNA模板,提高MSP退火温度能克服假阳性或假阴性结果的产生。结论 MSP检测甲基化具有高灵敏度的优点,适当提高退火温度能保证MSP结果的特异性。Objective To preliminarily assess the value of methylation-specilic polymerase chain reaction （MSP） for DNA methylation measurement. Methods Plasmids derived from hypermethylated clone and unmethylat- ed clone for SHH gene promoter were served as template to assess the sensitivity and specificity of MSP. Results MSP could detect low content of methylated DNA template. False positivity and false negativity could be overcomed by elevating annealing temperature of MSP. Conclusion MSP has the advantage of high sensitivity for methylated se quences. The specificity of MSP can be warranted by properly elevating annealing temperature.

关 键 词：甲基化特异性PCR 甲基化检测 敏感性 特异性 

分 类 号：R734.2[医药卫生—肿瘤]