Characterization of genomic single nucleotide polymorphism via colorimetric detection using a single gold nanoprobe |
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Authors: | Fábio Ferreira Carlos Orfeu Flores Gonçalo Doria Pedro Viana Baptista |
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Institution: | 1. Human Molecular Genetics Research Centre (CIGMH), Department of Life Sciences, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516 Caparica, Portugal;2. STAB VIDA–Investigação e Serviços em Ciências Biológicas, Madan Parque, 2825-182 Caparica, Portugal |
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Abstract: | Identification of specific nucleic acid sequences mediated by gold nanoparticles derivatized thiol-modified oligonucleotides (Au–nanoprobes) has been proven to be a useful tool in molecular diagnostics. Here, we demonstrate that, on optimization, detection may be simplified via the use of a single Au–nanoprobe to detect a single nucleotide polymorphism (SNP) in homo- or heterozygote condition. We validated this non-cross-linking approach through the analysis of 20 clinical samples using a single specific Au–nanoprobe for an SNP in the FTO (fat mass and obesity-associated) gene against direct DNA sequencing. Sensitivity, specificity, and limit of detection (LOD) were determined, and statistical differences were calculated by one-way analysis of variance (ANOVA) and a post hoc Tukey’s test to ascertain whether there were any differences between Au–nanoprobe genotyped groups. For the first time, we show that the use of a single Au–nanoprobe can detect SNP for each genetic status (wild type, heterozygous, or mutant) with high degrees of sensitivity (87.50%) and specificity (91.67%). |
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Keywords: | Gold nanoparticles Nanodiagnostics Non-cross-linking method Point of care FTO gene Genotyping |
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