title: Self-assembled biosensor with universal reporter and dual-quenchers for detection of unlabelled nucleic acids.
abstract: A novel biosensor with universal reporter and dual quenchers was developed for rapid, sensitive, selective, and inexpensive detection of unlabelled nucleic acids. The biosensor is based on a single-strand DNA stem-loop motif with an extended universal reporter-binding region, a G-base rich stem region, and a universal address-binding region. The self-assembly of these stem-loop probes with fluorescence labeled universal reporter and a universal address region conjugated to gold nanoparticles forms the basis of a biosensor for DNA or microRNA targets in solution. The introduction of dual quenchers (G-base quenching and gold surface plasmon resonance-induced quenching) significantly reduces the fluorescence background to as low as 12% of its original fluorescence intensity and hence enhances the detection limit to 0.01 picomoles without signal ampilication.
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, University of Nevada, Reno, NV 89557, USA. huang@medicine.nevada.edu.
authors.forename: Liming
authors.initials: L
authors.lastname: Huang
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, University of Nevada, Reno, NV 89557, USA. huang@medicine.nevada.edu.
authors.forename: Gyan H
authors.initials: GH
authors.lastname: Aryal
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, University of Nevada, Reno, NV 89557, USA. huang@medicine.nevada.edu.
authors.forename: Suk-Wah
authors.initials: SW
authors.lastname: Tam-Chang
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, University of Nevada, Reno, NV 89557, USA. huang@medicine.nevada.edu.
authors.forename: Nelson G
authors.initials: NG
authors.lastname: Publicover
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, University of Nevada, Reno, NV 89557, USA. huang@medicine.nevada.edu.
authors.forename: Kenneth W
authors.initials: KW
authors.lastname: Hunter
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D055029:Inverted Repeat Sequences; D053768:Metal Nanoparticles / Q000737:chemistry; D035683:MicroRNAs / Q000032:analysis* / Q000737:chemistry; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics; D013050:Spectrometry, Fluorescence
chemical_list: D035683:MicroRNAs; D015345:Oligonucleotide Probes; D006046:Gold; D004247:DNA