Journal
BIOSENSORS & BIOELECTRONICS
Volume 101, Issue -, Pages 29-36Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2017.10.011
Keywords
Dextrin gold nanoparticles; DNA; Salt; Stabilization; Genomic; Pathogen
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Funding
- C.S. Mott Predoctoral Fellowship in Sustainable Agriculture from Michigan State University
- Midland Research Institute for Value Chain Creation
- Michigan State University College of Natural Resources Undergraduate Research Program
- Michigan State University Rackham Foundation
- Michigan State University Project GREEEN [GR14-017]
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The interaction between gold nanoparticles (AuNPs) and nucleic acids has facilitated a variety of diagnostic applications, with further diversification of synthesis match bio-applications while reducing biotoxicity. However, DNA interactions with unique surface capping agents have not been fully defined. Using dextrin capped AuNPs (d-AuNPs), we have developed a novel unamplified genomic DNA (gDNA) nanosensor, exploiting dispersion and aggregation characteristics of d-AuNPs, in the presence of gDNA, for sequence-specific detection. We demonstrate that d-AuNPs are stable in a five-fold greater salt concentration than citrate-capped AuNPs and the d-AuNPs were stabilized by single stranded DNA probe (ssDNAp). However, in the elevated salt concentrations of the DNA detection assay, the target reactions were surprisingly further stabilized by the formation of a ssDNAp-target gDNA complex. The results presented herein lead us to propose a mechanism whereby genomic ssDNA secondary structure formation during ssDNAp-to-target gDNA binding enables d-AuNP stabilization in elevated ionic environments. Using the assay described herein, we were successful in detecting as little as 2.94 fM of pathogen DNA, and using crude extractions of a pathogen matrix, as few as 18 spores/mu L.
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