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Author:
Date:
2013-08-20
[Abstract] Detection of low copies of methylated DNA targets in clinical specimens is challenging. The quantitative Methylation-Specific PCR (qMSP) assays were designed to specifically amplify bisulphite-converted methylated DNA target sequences in the presence of an excess of unmethylated counterpart sequences. These qMSP assays are real-time PCR assays utilizing, sequence-specific primers and an intervening, also sequence specific, Taqman probe to cover an amplicon of approximately 100 bp in length. The use of Taqman probes bearing a minor groove binding (MGB) allow for the use of shorter probes and therefore facilitate design and significantly increases the analytical specificity of the reaction. In the context of the biomarker discovery program of the Liverpool Lung Project (LLP), ten gene ...
[摘要] 在临床标本中检测低拷贝的甲基化DNA靶是具有挑战性的。设计定量甲基化特异性PCR（qMSP）测定以在过量的未甲基化的对应物序列存在下特异性扩增亚硫酸氢盐转化的甲基化DNA靶序列。这些qMSP测定法是实时PCR测定，利用序列特异性引物和间插的，也是序列特异性的Taqman探针以覆盖长度约100bp的扩增子。使用具有小沟结合（MGB）的Taqman探针允许使用更短的探针，因此便于设计并显着增加反应的分析特异性。在利物浦肺项目（LLP）的生物标志物发现程序的上下文中，选择十个基因启动子。 qMSP测定法被开发，验证并用于筛选来自患有肺癌和具有非恶性肺疾病的年龄/性别匹配对照的患者的支气管洗液655（Nikolaidis等人，2012）。
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