Rapid KRAS Mutation Detection via Hybridization-Induced Aggregation of Microbeads

Analytical Chemistry
Hillary S SloaneJames P Landers

Abstract

Using hybridization-induced aggregation (HIA), a unique bead-based DNA detection technology scalable for a microchip platform, we describe a simplistic, low-cost method for rapid mutation testing. HIA utilizes a pair of sequence-specific oligonucleotide probes bound to magnetic microbeads. Hybridization to a target DNA strand tethers the beads together, inducing bead aggregation. By simply using the extent of bead aggregation as a measure of the hybridization efficiency, we avoid the need for additional labels and sophisticated analytical equipment. Through strategic manipulation of the assay design and experimental parameters, we use HIA to facilitate, for the first time, the detection of single base mutations in a gene segment and, specifically, the detection of activating KRAS mutations. Following the development and optimization of the assay, we apply it for KRAS mutation analysis of four human cancer cell lines. Ultimately, we present a proof-of-principle method for detecting any of the common KRAS mutations in a single-step, 2 min assay, using only one set of oligonucleotide probes, for a total analysis time of less than 10 min post-PCR. The assay is performed at room temperature and uses simple, inexpensive instrumentati...Continue Reading

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Citations

Jun 13, 2016·The Journal of Molecular Diagnostics : JMD·Hillary S SloaneKimberly A Kelly
Jun 9, 2016·Sensors·Stefan SchrittwieserJoerg Schotter
Oct 26, 2016·Analytical Chemistry·Kevin D ClarkJared L Anderson
Feb 29, 2020·Chemical Society Reviews·Kaixiang ZhangJinghong Li
Mar 8, 2019·ACS Sensors·Alireza Abi, Afsaneh Safavi

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