Data content for main Figures (1-6) in PLOS ONE paper "Nanopore-based Target Sequence Detection"

Abstract from the paper: The promise of portable diagnostic devices relies on three basic requirements: comparable sensitivity to established platforms, inexpensive manufacturing and cost of operations, and the ability to survive rugged field conditions. Solid state nanopores can meet all these requirements, but to achieve high manufacturing yields at low costs, assays must be tolerant to fabrication imperfections and to nanopore enlargement during operation. This paper presents a model for molecular engineering techniques that meets these goals with the aim of detecting target sequences within DNA. In contrast to methods that require precise geometries, we demonstrate detection using a range of pore geometries. As a result, our assay model tolerates any pore-forming method and in-situ pore enlargement. Using peptide nucleic acid (PNA) probes modified for conjugation with synthetic bulk-adding molecules, pores ranging 15-50 nm in diameter are shown to detect individual PNA-bound DNA. Detection of the CFTRDeltaF508 gene mutation, a codon deletion responsible for ~66% of all cystic fibrosis chromosomes, is demonstrated with a 26-36 nm pore size range by using a size-enhanced PNA probe. A mathematical framework for assessing the statistical significance of detection is also presented.

本论文摘要：便携式诊断设备的应用前景需满足三项基本要求：与成熟检测平台相当的灵敏度、低廉的制造成本与运营成本，以及可在严苛野外环境下稳定运行的能力。固态纳米孔（solid state nanopore）可满足上述全部要求，但为实现低成本下的高制造良率，检测分析方法需能够耐受制造过程中的缺陷，以及运行过程中纳米孔的孔径扩张。本文提出一种适配上述目标的分子工程技术模型，旨在检测DNA内的靶标序列。与依赖精准孔道几何结构的检测方法不同，本研究通过一系列不同孔道几何结构实现了检测。由此，本检测模型可兼容任意孔道制备方法，以及原位孔径扩张过程。通过与合成体积加成分子偶联修饰的肽核酸（peptide nucleic acid, PNA）探针，直径范围15~50 nm的纳米孔可实现对单个结合PNA的DNA分子的检测。本研究还利用体积增强型PNA探针，在孔径范围26~36 nm的纳米孔中实现了对囊性纤维化跨膜传导调节蛋白（CFTR）ΔF508基因突变的检测——该突变属于密码子缺失型，约占所有囊性纤维化染色体的66%。本文同时提出了用于评估检测结果统计学显著性的数学框架。