A novel DNA methylation biosensor by combination of isothermal amplification and lateral flow device
A novel DNA methylation biosensor by combination of isothermal amplification and lateral flow device
复制标题
等温扩增与侧流装置相结合的新型DNA甲基化生物传感器
DOI：
10.1016/j.snb.2021.129624
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发表时间：
2021-02-09
影响因子：
8.4
通讯作者：
Cui, Yali
中科院分区：
文献类型：
--
作者：
Liu, Xiaonan;Zhang, Jiaxing;Cui, Yali
DNA甲基化模式的异常已被证明是一个有前途的生物标志物，从临床角度对癌症的早期诊断，预后和治疗。在精准医学时代，目前的DNA甲基化鉴定技术不仅需要专业和耗时的操作，而且需要复杂的仪器和复杂的数据解释，这突出了开发直接和胜任的DNA甲基化检测策略的必要性。因此，本研究致力于建立一种新型的环介导等温扩增（LAMP）和侧流装置（LFD）相结合的生物传感器，用于从高度异质性的癌组织中检测微量DNA甲基化。为了解决基于核酸扩增的检测方法中灵敏度和特异性之间的天然矛盾，我们首次将生物素标记的内引物和地高辛标记的dUTP引入LAMP-LFD，使得该生物传感器在亚硫酸氢盐转化后35 min内在强干扰背景下识别低至150个拷贝或0.1%的目标变体。此外，在LFD中用作信号发生器的金磁性纳米颗粒能够通过视觉和磁性检测来解释结果。利用LAMP-LFD对miR-34 a启动子甲基化模式进行了研究，这表明该生物传感器可以很容易地适用于检测其他DNA甲基化生物标志物，作为癌症诊断的敏感筛选工具。
The aberration in DNA methylation pattern has been proved as a promising biomarker for early cancer diagnosis, prognosis and therapy from clinical perspective. In the era of precision medicine, current technologies for DNA methylation identification require not only professional and time-consuming operation but also sophisticated apparatus and complex data interpretation, which has highlighted the need to develop a straightforward and competent strategy for DNA methylation detection. Therefore, the present research is dedicated to establish a novel biosensor by integrating of loop-mediated isothermal amplification (LAMP) and lateral flow device (LFD) to identify trace of DNA methylation from highly heterogeneous cancerous specimen. In order to resolve the natural contradiction between sensitivity and specificity in nucleic acid amplification-based detection methods, we introduce the biotin labeled inner primer and digoxigenin labeled dUTP into LAMP-LFD for the first time, which allow this biosensor to identify as low as 150 copies or 0.1 % target variants under strong interferential background within 35 min after bisulfite conversion. Furthermore, the gold magnetic nanoparticle serving as a signal generator in LFD enables the result to be interpreted through both visual and magnetic detection. The promoter methylation pattern of miR-34a, a significant tumor suppressor gene, has been investigated using LAMP-LFD with cell lines and clinical samples, which indicate that this biosensor may be readily adapted for detection of other DNA methylation biomarkers as a sensitive screening tool for cancer diagnosis.