title: Characterization of genomic single nucleotide polymorphism via colorimetric detection using a single gold nanoprobe.
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%).
authors: 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; STAB VIDA-Investigação e Serviços em Ciências Biológicas, Madan Parque, 2825-182 Caparica, Portugal.
authors: Fábio Ferreira
authors: FF
authors: Carlos
authors: STAB VIDA-Investigação e Serviços em Ciências Biológicas, Madan Parque, 2825-182 Caparica, Portugal.
authors: Orfeu
authors: O
authors: Flores
authors: 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.
authors: Gonçalo
authors: G
authors: Doria
authors: 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. Electronic address: pmvb@fct.unl.pt.
authors: Pedro Viana
authors: PV
authors: Baptista
mesh_terms: D000071516:Alpha-Ketoglutarate-Dependent Dioxygenase FTO; D003124:Colorimetry / Q000379:methods; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D005260:Female; D060005:Genotyping Techniques / Q000379:methods*; D006046:Gold / Q000737:chemistry*; D006801:Humans; D008297:Male; D053768:Metal Nanoparticles / Q000737:chemistry*; D020641:Polymorphism, Single Nucleotide; D011506:Proteins / Q000235:genetics*
chemical_list: D015342:DNA Probes; D011506:Proteins; D006046:Gold; D000071516:Alpha-Ketoglutarate-Dependent Dioxygenase FTO; C526492:FTO protein, human
keywords: FTO gene; Genotyping; Gold nanoparticles; Nanodiagnostics; Non-cross-linking method; Point of care