학술논문
Label-free discrimination of single nucleotide changes in DNA by reflectometric interference Fourier transform spectroscopy.
Document Type
Academic Journal
Author
Makiyan F; Division of Nanobiotechnology, Department of Life Sciences Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran.; Rahimi F; Division of Nanobiotechnology, Department of Life Sciences Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran. Electronic address: rahimi.f@ut.ac.ir.; Hajati M; Division of Nanobiotechnology, Department of Life Sciences Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran.; Shafiekhani A; Physics Department, Alzahra University, Tehran, Iran; School of Physics, Institute for Research in Fundamental Sciences, Tehran, Iran.; Rezayan AH; Division of Nanobiotechnology, Department of Life Sciences Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran.; Ansari-Pour N; Division of Biotechnology, Department of Life Sciences Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran; Big Data Institute, Nuffield Department of Medicine, University of Oxford, Oxford, UK. Electronic address: n.ansaripour@ut.ac.ir.
Source
Publisher: Elsevier Country of Publication: Netherlands NLM ID: 9315133 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-4367 (Electronic) Linking ISSN: 09277765 NLM ISO Abbreviation: Colloids Surf B Biointerfaces Subsets: MEDLINE
Subject
Language
English
Abstract
Phenotypic variation - such as disease susceptibility and differential drug response - has a strong genetic component. Substantial effort has therefore been made to identify causal genomic variants explaining such variation among humans. Point mutations (PMs), which are single nucleotide changes in the genome, have been identified to be the most abundant form of causal genomic variants, making them useful, reliable diagnostic markers. Methods developed to genotype PMs have moved towards solid-phase assays, which not only show greater sensitivity and specificity, but also enable scalability and faster processing time. Most current assays are, however, based on fluorescent probes, which makes them relatively expensive. To develop a more cost-effective label-free genotyping method, we used a porous silicon (PSi) base as an efficient support for DNA biosensing and coupled it with reflectometric interference Fourier transform spectroscopy (RIFTS). To assess the versatility of this approach, we tested both a single nucleotide substitution in VKORC1 (-1639G > A; rs9923231) and a single nucleotide insertion in BRCA1 (5382insC; rs80357906). We demonstrate that the PSi-RIFTS method can efficiently detect both PM types with high sensitivity where hybridization of complementary DNA can be quantifiably differentiated from mismatch and non-complementary hybridization events. In addition, we show that the PSi base with immobilized DNA not only can be re-used to type further samples, but it also remains stable for 14 days, suggesting its potential for high-throughput applications.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
(Copyright © 2019 Elsevier B.V. All rights reserved.)