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A Unified Sensor Architecture for Isothermal Detection of Double‐Stranded DNA, Oligonucleotides, and Small Molecules
Author(s) -
Brown Carl W.,
Lakin Matthew R.,
FabryWood Aurora,
Horwitz Eli K.,
Baker Nicholas A.,
Stefanovic Darko,
Graves Steven W.
Publication year - 2015
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201402615
Subject(s) - oligonucleotide , dna , loop mediated isothermal amplification , double stranded , molecule , isothermal process , computational biology , chemistry , biophysics , nanotechnology , combinatorial chemistry , biological system , computer science , materials science , physics , biology , biochemistry , thermodynamics , organic chemistry
Pathogen detection is an important problem in many areas of medicine and agriculture, which can involve genomic or transcriptomic signatures or small‐molecule metabolites. We report a unified, DNA‐based sensor architecture capable of isothermal detection of double‐stranded DNA targets, single‐stranded oligonucleotides, and small molecules. Each sensor contains independent target detection and reporter modules, enabling rapid design. We detected gene variants on plasmids by using a straightforward isothermal denaturation protocol. The sensors were highly specific, even with a randomized DNA background. We achieved a limit of detection of ∼15 p M for single‐stranded targets and ∼5 n M for targets on denatured plasmids. By incorporating a blocked aptamer sequence, we also detected small molecules using the same sensor architecture. This work provides a starting point for multiplexed detection of multi‐strain pathogens, and disease states caused by genetic variants (e.g., sickle cell anemia).