RT Journal Article
T1 Oligonucleotide Sensor Based on Selective Capture of Upconversion Nanoparticles Triggered by Target Induced DNA Inter-Strand Ligand Reaction
A1 Méndez González, Diego
A1 Laurenti, Marco
A1 Latorre, Alfonso
A1 Somoza, Álvaro
A1 Vázquez, Ana
A1 Negredo, Ana Isabel
A1 López Cabarcos, Enrique
A1 Calderón, Oscar Gómez
A1 Melle Hernández, Sonia
A1 Rubio Retama, Jorge
AB We present a sensor that exploits the phenomenon of upconversion luminescence to detect the presence of specific sequences of small oligonucleotides like miRNAs among others. The sensor is based on NaYF4:Yb,Er@SiO2 nanoparticles functionalized with ssDNA that contain azide groups on the 3' ends. In the presence of a target sequence, inter-strand ligation is possible via click-reaction between one azide of the upconversion probe and a DBCO-ssDNA-biotin probe present in the solution. As result of this specific and selective process, biotin is covalently attached to the surface of the upconversion nanoparticles. The presence of biotin on the surface of the nanoparticles allows their selective capture on a streptavidin-coated support, giving a luminescent signal proportional to the amount of target present in the test samples. With the aim of studying the analytical properties of the sensor, total RNA samples were extracted from healthy mosquitoes and spiked-in with a specific target sequence at different concentrations. The result of these experiments revealed that the sensor was able to detect 10-17 moles (100 fM) of the target sequence in mixtures containing 100 ng of total RNA per well. Similar limit of detection was found for spiked human serum samples, demonstrating its suitability for detecting specific sequences of small oligonucleotides under real conditions. By contrast, under the presence of non-complementary sequences or sequences having mismatches, the luminescent signal was negligible or conspicuously reduced.
PB Amer Chemical Soc.
SN 1944-8244
YR 2017
FD 2017-03-23
LK https://hdl.handle.net/20.500.14352/17708
UL https://hdl.handle.net/20.500.14352/17708
LA eng
NO Es una versión postprint del artículo final.
NO Ministerio de Economía, Industria y Competitividad (MINECO)
NO Bill & Melinda Gates Foundation
NO Asociación Española Contra el Cáncer
NO Universidad Complutense-Santander
DS Docta Complutense
RD 27 jul 2024