| 甲基化特异性PCR的评价与优化 |
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| 引用本文： | 付祥胜,张亚历,李丽,肖岚月,邱野,周贤,张巍. 甲基化特异性PCR的评价与优化[J]. 检验医学与临床, 2012, 9(15): 1862-1863,1865 |
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| 作者姓名： | 付祥胜 张亚历 李丽 肖岚月 邱野 周贤 张巍 |
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| 作者单位： | 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 |
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| 基金项目： | 泸州医学院自然科学基金资助课题 |
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| 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.
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| 关 键 词： | 甲基化特异性PCR 甲基化检测 敏感性 特异性 |
The evaluation and optimization of methylation-specific polymerase chain reaction |
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| FU Xiang-sheng , ZHANG Ya-li , LI Li , XIAO Lan-yue , QIU Ye , ZHOU Xian , ZHANG Wei. The evaluation and optimization of methylation-specific polymerase chain reaction[J]. Laboratory Medicine and Clinic, 2012, 9(15): 1862-1863,1865 |
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| Authors: | FU Xiang-sheng ZHANG Ya-li LI Li XIAO Lan-yue QIU Ye ZHOU Xian ZHANG Wei |
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| Affiliation: | 1. Department of Digestive, Affiliated Hos- pital of Luzhou Medical College, Sichuan 646000, China ; 2. Institute for Digestive Medicine, Nan fang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, China) |
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| Abstract: | 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. |
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| Keywords: | methylation-specific polymerase chain reaction methylation measurement sensitivity spe-cificity |
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