A simple colorimetric detection of DNA methylation.
A simple colorimetric detection of DNA methylation.
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DOI：
10.1039/c2an35043b
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发表时间：
2012-04
期刊：
影响因子：
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通讯作者：
Chenchen Ge;Z. Fang;Junhua Chen;Jie Liu;Xuewen Lu;Lingwen Zeng
中科院分区：
文献类型：
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作者：
Chenchen Ge;Z. Fang;Junhua Chen;Jie Liu;Xuewen Lu;Lingwen Zeng
在这项工作中，我们描述了一个简单的比色法来检测DNA甲基化。合成并测试了具有含有甲基化胞嘧啶的小CpG区域（甲基化APC）的腺瘤性结肠息肉病（APC）。甲基化APC首先被抗5-甲基胞嘧啶单克隆抗体标记的磁性微球（MMPs）捕获和富集。然后加入与APC序列部分互补的探针，导致DNA双链体的形成。然后通过热变性释放微球捕获的探针，并将其添加到未修饰的金纳米颗粒（AuNP）溶液中。通过盐诱导聚集进行比色检测。检测限为80 fmol。用UV/维斯分光光度计通过记录AuNP溶液在520 nm处的吸光度进行半定量分析。该方法为DNA甲基化检测提供了一种简便、快速、定量的方法。
In this work, we describe a simple colorimetric method to detect DNA methylation. Adenomatous polyposis coli (APC) with a small CpG region containing methylated cytosine (methylated APC) was synthesized and tested. Methylated APC was first captured and enriched by anti-5-methylcytosine monoclonal antibody conjugated magnetic microspheres (MMPs). Then a probe partly complementary to the APC sequence was added, resulting in the formation of DNA duplexes. The microsphere-captured probe was then released by heat denaturation and added into unmodified gold nanoparticle (AuNP) solution. Colorimetric detection was performed by salt-induced aggregation. The limit of detection is 80 fmol. Semi-quantitative analysis was done with a UV/Vis spectrophotometer by recording the absorbance of AuNP solution at 520 nm. Thus, this method provides a simple, rapid and quantitative tool for DNA methylation detection.