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    • 2. 发明申请
    • Method for the quantification of methylated DNA
    • 甲基化DNA定量的方法
    • US20050287553A1
    • 2005-12-29
    • US11100779
    • 2005-04-06
    • David GuetigDirk HabighorstAntje KluthArmin SchmittMatthias SchusterIna Schwope
    • David GuetigDirk HabighorstAntje KluthArmin SchmittMatthias SchusterIna Schwope
    • C12Q1/68G06F19/00C12P19/34
    • C12Q1/6823C12Q2565/101C12Q2535/131C12Q2545/114C12Q2523/125
    • Particular aspects of the present invention provide a method for quantification of two different variations of a DNA sequence. Particularly, the invention relates to a quantification of methylated DNA, and for this purpose, the test DNA is converted so that cytosine is converted to uracil, while 5-methylcytosine remains unchanged. The converted DNA is amplified by means of a real-time PCR, wherein two labeled real-time probe types are utilized: one specific for methylated DNA; and one for unmethylated DNA. Preferably, the degree of methylation of the test DNA is calculated from the ratio of the signal intensities of the probes or from the Ct values. The inventive methods have substantial utility for diagnosis and prognosis of cancer and other disorders associated with altered or characteristic DNA methylation status, as well as having substantial utility for analysis of SNPs, allelic expression, and prediction of drug response, drug interactions, among other uses.
    • 本发明的特定方面提供了用于定量DNA序列的两个不同变体的方法。 特别地,本发明涉及甲基化DNA的定量,为此目的,转化测试DNA使得胞嘧啶转化为尿嘧啶,而5-甲基胞嘧啶保持不变。 通过实时PCR扩增转化的DNA,其中使用两种标记的实时探针类型:一种特异性用于甲基化DNA; 一个用于未甲基化的DNA。 优选地,根据探针的信号强度与Ct值的比值来计算测试DNA的甲基化程度。 本发明的方法具有用于诊断和预测与改变或特征性DNA甲基化状态相关的癌症和其他病症的实质性用途,并且具有用于分析SNP,等位基因表达和药物反应预测,药物相互作用以及其它用途的实质性用途 。