Many diseases are defined by patterns of DNA methylation which result in aberrant gene expression. We present a rapid assay based upon resistive pulse sensing, RPS, to characterize sequence specific DNA methylation sites in genomic DNA. We modify the surface of superparamagnetic beads, SPBs, with DNA (capture probe). The particles are added to solution where they bind to and extract sequence specific DNA (target DNA). The target loaded SPBs are then incubated with antibodies which bind to the methylation sites, and the velocity of the SPBs through the nanopore reveals the number and location of the epigenetic markers within the target. The approach is capable of distinguishing between different methylation sites within a DNA promoter region. Crucially the approach is not dependent on accurate sequencing of assayed DNA, with genomic regions targeted through complementary probes. As such the number of stages and reagents costs are low and the assay is complete in under 60 min which includes the incubation and run times. The format also allows simultaneous quantification of number of copies of methylated DNA, and we illustrate this with a dose response curve.

中文翻译：

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通过电阻脉冲传感快速评估位点特异性DNA甲基化

DNA甲基化的模式可导致基因表达异常，从而定义了许多疾病。我们提出了一种基于电阻性脉冲感应RPS的快速测定法，以表征基因组DNA中的序列特异性DNA甲基化位点。我们用DNA（捕获探针）修饰超顺磁珠SPB的表面。将颗粒添加到溶液中，在此与颗粒结合并提取序列特异性DNA（目标DNA）。然后将装有靶标的SPB与结合到甲基化位点的抗体一起孵育，SPB通过纳米孔的速度揭示了表观遗传标记在靶标内的数量和位置。该方法能够区分DNA启动子区域内的不同甲基化位点。至关重要的是，该方法不依赖于测定的DNA的准确测序，通过互补探针靶向的基因组区域。因此，阶段数和试剂成本低，并且测定可在60分钟内完成，包括孵育和运行时间。该格式还允许同时量化甲基化DNA的拷贝数，我们用剂量反应曲线说明了这一点。