通过单分子动力指纹识别对富含甲基CpG的癌症生物标志物的超敏感无放大量化
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Updated: Jun 16, 2026

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Targeted DNA Methylation Analysis by Next-generation Sequencing
Published on: February 24, 2015
37.1K
Liuhan Dai1,2, Alexander Johnson-Buck1, Peter W Laird3
1Single Molecule Analysis Group, Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, USA.
bioRxiv : the preprint server for biology
|April 22, 2024
概括
我们开发了一种新的,无放大方法,以准确量化DNA甲基化,这是一个关键的表观遗传标记. 这种技术可以在液体活检中提升甲基化癌症DNA生物标志物的早期检测.
科学领域:
- 表观遗传学 在表观遗传学中,表观遗传学是指表观遗传学.
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 5甲基细胞因是重要的表观遗传标记物,具有疾病生物标记物发展的潜力.
- 当前的DNA甲基化量化方法,如双硫酸PCR,容易产生放大偏差.
- 需要敏感和准确的无放大DNA甲基化试验.
研究的目的:
- 开发一种无放大试验,用于量化DNA甲基化.
- 评估新型试验的敏感性和特异性.
- 为了证明测试在复杂的基因组DNA矩阵中的实用性.
主要方法:
- 结合双硫酸盐转换与单分子动力指纹.
- 开发了一种针对BCAT1促进体的无放大试验.
- 在复杂的基因组DNA样本上验证了测量.
主要成果:
- 该试验表现出高特异性 (99.9999%) 和低检测极限 (<1 fM).
- 在BCAT1促进体中成功区分了低百分比 (2-5%) 的DNA甲基化.
- 在全血DNA矩阵中可靠量化甲基化.
结论:
- 无放大,单分子量化方法是可行的和敏感的.
- 这种方法比检测甲基化DNA生物标记物的现有技术提供了改进.
- 该试验对使用液体活检早期发现癌症具有前途.
相关概念视频
DNA Microarrays
Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
Real Time RT-PCR
Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
The real-time quantification of the number of amplified products is...

