2024-03-29T12:50:39Zhttp://repisalud.isciii.es/oai/requestoai:repisalud.isciii.es:20.500.12105/76802023-04-19T12:34:54Zcom_20.500.12105_2060com_20.500.12105_2052com_20.500.12105_2051col_20.500.12105_2061
00925njm 22002777a 4500
dc
Mendez-Gonzalez, Diego
author
Laurenti, Marco
author
Latorre, Alfonso
author
Somoza, Alvaro
author
Vazquez, Ana
author
Negredo, Anabel
author
López-Cabarcos, Enrique
author
Calderón, Oscar G.
author
Melle, Sonia
author
Rubio-Retama, Jorge
author
2017-03
We present a sensor that exploits the phenomenon of upconversion luminescence to detect the presence of specific sequences of small oligonucleotides such as 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, interstrand ligation is possible via the click-reaction between one azide of the upconversion probe and a DBCO-ssDNA-biotin probe present in the solution. As a 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 strands 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 were 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 per well (100 fM) of the target sequence in mixtures containing 100 ng of total RNA per well. A similar limit of detection was found for spiked human serum samples, demonstrating the suitability of the sensor for detecting specific sequences of small oligonucleotides under real conditions. In contrast, in the presence of noncomplementary sequences or sequences having mismatches, the luminescent signal was negligible or conspicuously reduced.
ACS Appl Mater Interfaces. 2017 Apr 12;9(14):12272-12281
1944-8244
http://hdl.handle.net/20.500.12105/7680
28332400
10.1021/acsami.7b00575
1944-8252
ACS Applied Materials & Interfaces
DNA
Interstrand ligation
Nanoparticles
Sensor
Upconversion
Oligonucleotide Sensor Based on Selective Capture of Upconversion Nanoparticles Triggered by Target-Induced DNA Interstrand Ligand Reaction