Shepherd Britney A, Tanjil Md Rubayat-E, Jeong Yunjo, Baloğlu Bilgenur, Liao Jingqiu, Wang Michael Cai
Department of Medical Engineering, University of South Florida, 4202 E. Fowler Avenue, Tampa, Florida 33620 USA.
Department of Mechanical Engineering, University of South Florida, 4202 E. Fowler Avenue, Tampa, Florida 33620 USA.
MRS Adv. 2020;5(56):2889-2906. doi: 10.1557/adv.2020.402. Epub 2020 Dec 1.
State-of-the-art nanopore sequencing enables rapid and real-time identification of novel pathogens, which has wide application in various research areas and is an emerging diagnostic tool for infectious diseases including COVID-19. Nanopore translocation enables de novo sequencing with long reads (> 10 kb) of novel genomes, which has advantages over existing short-read sequencing technologies. Biological nanopore sequencing has already achieved success as a technology platform but it is sensitive to empirical factors such as pH and temperature. Alternatively, ångström- and nano-scale solid-state nanopores, especially those based on two-dimensional (2D) membranes, are promising next-generation technologies as they can surpass biological nanopores in the variety of membrane materials, ease of defining pore morphology, higher nucleotide detection sensitivity, and facilitation of novel and hybrid sequencing modalities. Since the discovery of graphene, atomically-thin 2D materials have shown immense potential for the fabrication of nanopores with well-defined geometry, rendering them viable candidates for nanopore sequencing membranes. Here, we review recent progress and future development trends of 2D materials and their ångström- and nano-scale pore-based nucleic acid (NA) sequencing including fabrication techniques and current and emerging sequencing modalities. In addition, we discuss the current challenges of translocation-based nanopore sequencing and provide an outlook on promising future research directions.
最先进的纳米孔测序技术能够快速实时鉴定新型病原体，在各个研究领域有着广泛应用，并且是包括新冠病毒肺炎在内的传染病的一种新兴诊断工具。纳米孔易位能够对新基因组进行从头测序，产生长读长（>10 kb），这比现有的短读长测序技术具有优势。生物纳米孔测序作为一种技术平台已经取得成功，但它对诸如pH值和温度等经验因素敏感。另外，埃米级和纳米级固态纳米孔，尤其是基于二维（2D）膜的纳米孔，作为下一代技术很有前景，因为它们在膜材料种类、易于定义孔形态、更高的核苷酸检测灵敏度以及促进新型和混合测序模式等方面能够超越生物纳米孔。自石墨烯被发现以来，原子级薄的二维材料在制造具有明确几何形状的纳米孔方面显示出巨大潜力，使其成为纳米孔测序膜的可行候选材料。在此，我们综述二维材料及其基于埃米级和纳米级孔的核酸（NA）测序的最新进展和未来发展趋势，包括制造技术以及当前和新兴的测序模式。此外，我们讨论基于易位的纳米孔测序当前面临的挑战，并对未来有前景的研究方向进行展望。
