Oligonucleotide Sensor Based on Selective Capture of Upconversion Nanoparticles Triggered by Target-Induced DNA Interstrand Ligand Reaction.
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@SiO2nanoparticles 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 10-17moles 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.
Department of Physical Chemistry II, Faculty of Pharmacy, Complutense University of Madrid , 28040 Madrid, Spain.
Diego
D
Mendez-Gonzalez
Department of Physical Chemistry II, Faculty of Pharmacy, Complutense University of Madrid , 28040 Madrid, Spain.
Marco
M
Laurenti
Nanobiotecnología (IMDEA-Nanociencia), Unidad Asociada al Centro Nacional de Biotecnología (CSIC) , 28049 Madrid, Spain.
Alfonso
A
Latorre
Nanobiotecnología (IMDEA-Nanociencia), Unidad Asociada al Centro Nacional de Biotecnología (CSIC) , 28049 Madrid, Spain.
Alvaro
http://orcid.org/0000-0001-9873-435X
A
Somoza
Laboratorio de Arbovirus, Centro Nacional de Microbiología-Instituto de Salud Carlos III , Majadahonda, 28220 Madrid, Spain.
Ana
A
Vazquez
Laboratorio de Arbovirus, Centro Nacional de Microbiología-Instituto de Salud Carlos III , Majadahonda, 28220 Madrid, Spain.
Ana Isabel
AI
Negredo
Department of Physical Chemistry II, Faculty of Pharmacy, Complutense University of Madrid , 28040 Madrid, Spain.
Enrique
E
López-Cabarcos
Faculty of Optics and Optometry, Complutense University of Madrid , Arcos de Jalón 118, 28037 Madrid, Spain.
Oscar G
OG
Calderón
Faculty of Optics and Optometry, Complutense University of Madrid , Arcos de Jalón 118, 28037 Madrid, Spain.
Sonia
S
Melle
Department of Physical Chemistry II, Faculty of Pharmacy, Complutense University of Madrid , 28040 Madrid, Spain.
Jorge
http://orcid.org/0000-0002-1785-5844
J
Rubio-Retama
D004247:DNA; D006801:Humans; D049449:Luminescence; D053758:Nanoparticles; D009841:Oligonucleotides; D012822:Silicon Dioxide
D009841:Oligonucleotides; D012822:Silicon Dioxide; D004247:DNA
DNA; interstrand ligation; nanoparticles; sensor; upconversion