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PLoS One. 2016 May 5;11(5):e0154426. doi: 10.1371/journal.pone.0154426. eCollection 2016.

Nanopore-Based Target Sequence Detection.

Author information

1
Two Pore Guys Inc., Santa Cruz, CA, United States of America.
2
Department of Physics, University of Ottawa, Ontario, Canada.
3
Baskin School of Engineering, University of California Santa Cruz, Santa Cruz, CA, United States of America.

Abstract

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 CFTRΔF508 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.

PMID:
27149679
PMCID:
PMC4858282
DOI:
10.1371/journal.pone.0154426
[Indexed for MEDLINE]
Free PMC Article

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