title: A unified sensor architecture for isothermal detection of double-stranded DNA, oligonucleotides, and small molecules.
abstract: 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 pM for single-stranded targets and ∼5 nM 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).
authors: Center for Biomedical Engineering, University of New Mexico, Albuquerque, NM 87131 (USA).
authors: Carl W
authors: CW
authors: Brown
authors: Matthew R
authors: MR
authors: Lakin
authors: Aurora
authors: A
authors: Fabry-Wood
authors: Eli K
authors: EK
authors: Horwitz
authors: Nicholas A
authors: NA
authors: Baker
authors: Darko
authors: D
authors: Stefanovic
authors: Steven W
authors: SW
authors: Graves
mesh_terms: D015374:Biosensing Techniques; D004247:DNA / Q000032:analysis* / Q000235:genetics; D015342:DNA Probes / Q000032:analysis* / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis; D009691:Nucleic Acid Denaturation; D009841:Oligonucleotides / Q000032:analysis* / Q000235:genetics; D013696:Temperature
chemical_list: D015342:DNA Probes; D009841:Oligonucleotides; D049452:Green Fluorescent Proteins; D004247:DNA
keywords: DNAzymes; amplification; deoxyribozymes; pathogen detection; strand displacement