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Methylation-Sensitive Loop-Mediated Isothermal Amplification (LAMP): Nucleic Acid Methylation Detection through LAMP with Mobile Fluorescence Readout
ACS Sensors ( IF 8.2 ) Pub Date : 2021-09-01 , DOI: 10.1021/acssensors.1c00902 Jacob Amos Hambalek 1 , Janay Elise Kong 1 , Calvin Brown 2 , Hector Enrique Munoz 1 , Thomas Horn 1 , Michael Bogumil 1 , Eleni Quick 1 , Aydogan Ozcan 2 , Dino Di Carlo 1, 3
ACS Sensors ( IF 8.2 ) Pub Date : 2021-09-01 , DOI: 10.1021/acssensors.1c00902 Jacob Amos Hambalek 1 , Janay Elise Kong 1 , Calvin Brown 2 , Hector Enrique Munoz 1 , Thomas Horn 1 , Michael Bogumil 1 , Eleni Quick 1 , Aydogan Ozcan 2 , Dino Di Carlo 1, 3
Affiliation
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The emergence of epigenetic gene regulation and its role in disease have motivated a growing field of epigenetic diagnostics for risk assessment and screening. In particular, irregular cytosine DNA base methylation has been implicated in several diseases, yet the methods for detecting these epigenetic marks are limited to lengthy protocols requiring bulky and costly equipment. We demonstrate a simple workflow for detecting methylated CpG dinucleotides in synthetic and genomic DNA samples using methylation-sensitive restriction enzyme digestion followed by loop-mediated isothermal amplification. We additionally demonstrate a cost-effective mobile fluorescence reader comprising a light-emitting diode bundle, a mirror, and optical fibers to transduce fluorescence signals associated with DNA amplification. The workflow can be performed in approximately 1 h, requiring only a simple heat source, and can therefore provide a foundation for distributable point-of-care testing of DNA methylation levels.
中文翻译：
甲基化敏感环路介导的等温扩增 (LAMP)：通过 LAMP 检测核酸甲基化并带有移动荧光读数
表观遗传基因调控的出现及其在疾病中的作用推动了用于风险评估和筛查的表观遗传诊断领域的发展。特别是，不规则的胞嘧啶 DNA 碱基甲基化与多种疾病有关，但检测这些表观遗传标记的方法仅限于需要庞大且昂贵设备的冗长协议。我们展示了一个简单的工作流程，用于检测合成和基因组 DNA 样品中的甲基化 CpG 二核苷酸，使用甲基化敏感的限制酶消化，然后是环介导的等温扩增。我们还展示了一种具有成本效益的移动荧光读取器，包括发光二极管束、镜子和光纤，用于转导与 DNA 扩增相关的荧光信号。
更新日期：2021-09-24
中文翻译：
甲基化敏感环路介导的等温扩增 (LAMP)：通过 LAMP 检测核酸甲基化并带有移动荧光读数
表观遗传基因调控的出现及其在疾病中的作用推动了用于风险评估和筛查的表观遗传诊断领域的发展。特别是，不规则的胞嘧啶 DNA 碱基甲基化与多种疾病有关，但检测这些表观遗传标记的方法仅限于需要庞大且昂贵设备的冗长协议。我们展示了一个简单的工作流程，用于检测合成和基因组 DNA 样品中的甲基化 CpG 二核苷酸，使用甲基化敏感的限制酶消化，然后是环介导的等温扩增。我们还展示了一种具有成本效益的移动荧光读取器，包括发光二极管束、镜子和光纤，用于转导与 DNA 扩增相关的荧光信号。
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