期刊
BIOSENSORS & BIOELECTRONICS
卷 130, 期 -, 页码 236-244出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2019.01.045
关键词
Localized surface plasmon resonance (LSPR); Liquid biopsy; Nanosensor; Circulating tumor DNA; Point mutation; Nanorods; Surface conjugation
类别
资金
- National Institutes of Health (NIH) Director's Transformative Research Award (United States) [R01HL137157]
- Norris Cotton Cancer Center Developmental Funds Pilot Projects (United States)
- Thayer School of Engineering PhD Innovation Program (United States)
Here we present a gold nanorod-based platform for the sequence-specific detection of circulating tumor DNA (ctDNA) point mutations without the need for amplification or fluorescence labeling. Peptide nucleic acid probes complimentary to the G12V mutation in the KRAS gene were conjugated to gold nanorods, and the localized surface plasmon resonance absorbance through the sample was measured after exposure to synthetic ctDNA at various concentrations. Each step of the reaction was thoroughly controlled, starting from reagent concentrations and including conjugation, sonication, and incubation time. The platform was evaluated in both buffer and spiked healthy patient serum, demonstrating a linear working range below 125 nanograms of ctDNA per milliliter solution, and an effective limit of detection of 2 nanograms of ctDNA per milliliter. A clear distinction between mutant and wild type synthetic ctDNA was also found using this platform. In order to improve upon the selectivity of the sensor, a DNA hybridization simulation was performed to understand how the addition of mutations to the peptide nucleic acid probe could enhance the selectivity for capture of mutant over wild type sequences. The top candidate from the simulations, which had an additional mutation two base pairs away from the mutation of interest, had a significant impact on the selectivity between mutant and wild type capture. This paper provides a framework for sequence-specific capture of ctDNA, and a method of improving selectivity for desired point mutations through careful probe design.
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