DNA lesions such as metholcytosine(mC), 8-OXO-guanine (OG), inosine (I), etc. could cause genetic diseases. Identification of the varieties of lesion bases are usually beyond the capability of conventional DNA sequencing which is mainly designed to discriminate four bases only. Therefore, lesion detection remains a challenge due to massive varieties and less distinguishable readouts for structural variations at the molecular level. Moreover, standard amplification and labeling hardly work in DNA lesion detection. Herein, we designed a single molecule interface from the mutant aerolysin (K238Q), whose sensing region shows high compatibility to capture and then directly convert a minor lesion into distinguishable electrochemical readouts. Compared with previous single molecule sensing interfaces, the temporal resolution of the K238Q aerolysin nanopore is enhanced by two orders, which has the best sensing performance in all reported aerolysin nanopores. In this work, the novel K238Q could discriminate directly at least three types of lesions (mC, OG, I) without labeling and quantify modification sites under the mixed heterocomposition conditions of the oligonucleotide. Such a nanopore electrochemistry approach could be further applied to diagnose genetic diseases at high sensitivity.

中文翻译：

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使用气溶素单分子界面直接定量寡核苷酸中受损的核苷酸。

诸如胞嘧啶（mC），8-OXO-鸟嘌呤（OG），肌苷（I）等DNA损伤可能引起遗传疾病。病变碱基种类的鉴定通常超出了常规DNA测序的能力，常规DNA测序的主要目的是仅区分四个碱基。因此，由于种类繁多，并且在分子水平上对于结构变化的读数难以区分，病变检测仍然是一个挑战。此外，标准扩增和标记在DNA病变检测中几乎不起作用。在这里，我们从突变的溶血素（K238Q）设计了一个单分子界面，其传感区域显示出高的相容性，可以捕获然后直接将较小的病变转化为可区分的电化学读数。与以前的单分子传感接口相比，K238Q溶血素纳米孔的时间分辨率提高了两个数量级，在所有报道的溶血素纳米孔中具有最佳的传感性能。在这项工作中，新型K238Q可以直接区分至少三种类型的病变（mC，OG，I），而无需在寡核苷酸的混合杂合条件下进行标记和量化修饰位点。这种纳米孔电化学方法可以进一步应用于以高灵敏度诊断遗传疾病。