Optomagnetic Detection of MicroRNA Based on Duplex-Specific Nuclease-Assisted Target Recycling and Multilayer Core-Satellite Magnetic Superstructures.
Superstructural assembly of magnetic nanoparticles enables approaches to biosensing by combining specially tailored properties of superstructures and the particular advantages associated with a magnetic or optomagnetic read-out such as low background signal, easy manipulation, cost-efficiency, and potential for bioresponsive multiplexing. Herein, we demonstrate a sensitive and rapid miRNA detection method based on optomagnetic read-out, duplex-specific nuclease (DSN)-assisted target recycling, and the use of multilayer core-satellite magnetic superstructures. Triggered by the presence of target miRNA and DSN-assisted target recycling, the core-satellite magnetic superstructures release their "satellites" to the suspension, which subsequently can be quantified accurately in a low-cost and user-friendly optomagnetic setup. Target miRNAs are preserved in the cleaving reaction and can thereby trigger more cleavage and release of "satellites". For singleplex detection of let-7b, a linear detection range between 10 fM and 10 nM was observed, and a detection limit of 4.8 fM was obtained within a total assay time of 70 min. Multiplexing was achieved by releasing nanoparticles of different sizes in the presence of different miRNAs. The proposed method also has the advantages of single-nucleotide mismatch discrimination and the ability of quantification in a clinical sample matrix, thus holding great promise for miRNA routine multiplex diagnostics.
Department of Engineering Sciences, The Ångström Laboratory, Uppsala University , Box 534, SE-751 21 Uppsala, Sweden.
Bo
0000-0002-5249-4415
B
Tian
Department of Immunology, Genetics, and Pathology, The Rudbeck Laboratory, Uppsala University , SE-751 85 Uppsala, Sweden.
Jing
J
Ma
Department of Engineering Sciences, The Ångström Laboratory, Uppsala University , Box 534, SE-751 21 Uppsala, Sweden.
Zhen
Z
Qiu
Department of Engineering Sciences, The Ångström Laboratory, Uppsala University , Box 534, SE-751 21 Uppsala, Sweden.
Teresa
T
Zardán Gómez de la Torre
BluSense Diagnostics , Fruebjergvej 3, DK-2100 Copenhagen, Denmark.
Marco
M
Donolato
Department of Micro- and Nanotechnology, Technical University of Denmark, DTU Nanotech , Building 345B, DK-2800 Kongens Lyngby, Denmark.
Mikkel Fougt
MF
Hansen
Department of Engineering Sciences, The Ångström Laboratory, Uppsala University , Box 534, SE-751 21 Uppsala, Sweden.
Peter
P
Svedlindh
Department of Engineering Sciences, The Ångström Laboratory, Uppsala University , Box 534, SE-751 21 Uppsala, Sweden.
Mattias
0000-0003-0648-3130
M
Strömberg
D015374:Biosensing Techniques; D006801:Humans; D060328:Magnetic Phenomena; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D035683:MicroRNAs / Q000032:analysis*; D010316:Particle Size; D014407:Tumor Cells, Cultured
D058185:Magnetite Nanoparticles; D035683:MicroRNAs
core-satellite superstructures; duplex-specific nuclease; magnetic nanoparticles; miRNA detection; optomagnetic bioassay