Combination of methylated-DNA precipitation and methylation-sensitive restriction enzymes (COMPARE-MS) for the rapid, sensitive and quantitative detection of DNA methylation.
Combination of methylated-DNA precipitation and methylation-sensitive restriction enzymes (COMPARE-MS) for the rapid, sensitive and quantitative detection of DNA methylation.
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DOI：
10.1093/nar/gnj022
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发表时间：
2006-02-09
影响因子：
14.9
通讯作者：
Nelson WG
中科院分区：
文献类型：
--
作者：
Yegnasubramanian S;Lin X;Haffner MC;DeMarzo AM;Nelson WG
CpG岛超甲基化（CGI）是一种几乎普遍存在的肿瘤体细胞基因组改变。快速而敏感地检测DNA超甲基化将有助于癌症诊断和危险分层。我们提出了一种新的技术，称为COMPARE-MS，可以快速，定量检测CGI超甲基化，同时在数百个样品中具有高灵敏度和特异性。为了定量CGI超甲基化，COMPARE-MS使用DNA的实时PCR，该DNA首先被甲基化敏感的限制性内切酶消化，然后被固定在磁性固体基质上的甲基结合结构域多肽沉淀。我们表明，COMPARE-MS可以检测到5个基因组当量的甲基化CGIs在1000至10 000倍过量的未甲基化DNA。COMPARE-MS用于快速定量>155个前列腺组织（包括良性和恶性前列腺样本以及前列腺细胞系）中多个CGI处的超甲基化。该分析表明，通过COMPARE-MS测定的GST 1、MDR 1和PTGS 2 CGI超甲基化可以区分恶性和良性前列腺，敏感性>95%，特异性接近100%。这项新技术可以显着提高我们检测CGI超甲基化的能力。
Hypermethylation of CpG island (CGI) sequences is a nearly universal somatic genome alteration in cancer. Rapid and sensitive detection of DNA hypermethylation would aid in cancer diagnosis and risk stratification. We present a novel technique, called COMPARE-MS, that can rapidly and quantitatively detect CGI hypermethylation with high sensitivity and specificity in hundreds of samples simultaneously. To quantitate CGI hypermethylation, COMPARE-MS uses real-time PCR of DNA that was first digested by methylation-sensitive restriction enzymes and then precipitated by methyl-binding domain polypeptides immobilized on a magnetic solid matrix. We show that COMPARE-MS could detect five genome equivalents of methylated CGIs in a 1000- to 10 000-fold excess of unmethylated DNA. COMPARE-MS was used to rapidly quantitate hypermethylation at multiple CGIs in >155 prostate tissues, including benign and malignant prostate specimens, and prostate cell lines. This analysis showed that GSTP1, MDR1 and PTGS2 CGI hypermethylation as determined by COMPARE-MS could differentiate between malignant and benign prostate with sensitivities >95% and specificities approaching 100%. This novel technology could significantly improve our ability to detect CGI hypermethylation.
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DOI：
10.1073/pnas.93.18.9821
发表时间：
1996-09-03
影响因子：
11.1
作者：
Herman, JG;Graff, JR;Baylin, SB
通讯作者：
Baylin, SB
影响因子：
78.5
作者：
Issa, JP
通讯作者：
Issa, JP
DOI：
10.1073/pnas.77.11.6463
发表时间：
1980-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
POLLACK, Y;STEIN, R;CEDAR, H
通讯作者：
CEDAR, H
影响因子：
14.9
作者：
Cottrell, SE;Distler, J;Berlin, K
通讯作者：
Berlin, K
影响因子：
4.8
作者：
Bakker, J;Lin, XH;Nelson, WG
通讯作者：
Nelson, WG