高分辨率熔体
甲基化
CpG站点
DNA甲基化
表观遗传学
熔化曲线分析
计算生物学
热分析
心房颤动
生物
化学
生物信息学
材料科学
基因
实时聚合酶链反应
遗传学
内科学
基因表达
医学
聚合酶链反应
热的
物理
气象学
作者
Yan Yu,Sheng Wang,Yang Luo,Chang Gu,Xin Shi,Feng Shen
出处
期刊:ACS Sensors
[American Chemical Society]
日期:2023-08-17
卷期号:8 (9): 3595-3603
被引量:4
标识
DOI:10.1021/acssensors.3c01309
摘要
Methylation is an essential epigenetic modification involved in regulating gene expression and maintaining genome stability. Methylation patterns can be heterogeneous, exhibiting variations in both level and density. However, current methods of methylation analysis, including sequencing, methylation-specific PCR, and high-resolution melting curve analysis (HRM), face limitations of high cost, time-consuming workflows, and the difficulty of both accurate heterogeneity analysis and precise quantification. Here, a droplet array SlipChip-based (da-SlipChip-based) digital melting curve analysis (MCA) method was developed for the accurate quantification of both methylation level (ratio of methylated molecules to total molecules) and methylation density (ratio of methylated CpG sites to total CpG sites). The SlipChip-based digital MCA system supplements an in situ thermal cycler with a fluorescence imaging module for real-time MCA. The da-SlipChip can generate 10,656 droplets of 1 nL each, which can be separated into four lanes, enabling the simultaneous analysis of four samples. This method's clinical application was demonstrated by analyzing samples from ten healthy individuals and twenty patients with atrial fibrillation (AF), the most common arrhythmia. This method can distinguish healthy individuals from those with AF of both the paroxysmal and persistent types. It also holds potential for broader application in various research and clinical settings requiring methylation analysis.
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