title: Multiplexed microRNA detection using lanthanide-labeled DNA probes and laser ablation inductively coupled plasma mass spectrometry.
abstract: In the past decade, microRNAs (miRNAs) have drawn increasing attention due to their role in regulation of gene expression. Especially, their potential as biomarkers in disease diagnostics has motivated miRNA research, including the development of simple, accurate, and sensitive detection methods. The narrow size range of miRNAs (20-24 nucleotides) combined with the chemical properties of conventional reporter tags has hampered the development of multiplexed miRNA assays. In this study, we have used lanthanide-labeled DNA probes for the detection of miRNAs on membranes using laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS). Three miRNAs from Arabidopsis thaliana were analyzed simultaneously with high specificity, and the sensitivity of the method was comparable to radioactive detection (low femtomol range). The perspective of the developed method is highly multiplexed and quantitative miRNA analysis with high specificity and sensitivity.
authors.affiliation: Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen , Thorvaldsensvej 40, DK-1871 Frederiksberg C, Denmark.
authors.forename: Thomas Christian
authors.initials: TC
authors.lastname: de Bang
authors.forename: Pratik
authors.initials: P
authors.lastname: Shah
authors.forename: Seok Keun
authors.initials: SK
authors.lastname: Cho
authors.forename: Seong Wook
authors.initials: SW
authors.lastname: Yang
authors.forename: Søren
authors.initials: S
authors.lastname: Husted
mesh_terms: D017360:Arabidopsis / Q000235:genetics; D015152:Blotting, Northern / Q000295:instrumentation; D015342:DNA Probes / Q000737:chemistry*; D028581:Lanthanoid Series Elements / Q000737:chemistry*; D053685:Laser Therapy; D013058:Mass Spectrometry / Q000379:methods*; D008567:Membranes, Artificial; D035683:MicroRNAs / Q000032:analysis*; D010759:Phosphorus Isotopes; D018749:RNA, Plant / Q000032:analysis; D012680:Sensitivity and Specificity
chemical_list: D015342:DNA Probes; D028581:Lanthanoid Series Elements; C000600917:MIRN160 microRNA, Arabidopsis; C580969:MIRN166 microRNA, Arabidopsis; D008567:Membranes, Artificial; D035683:MicroRNAs; C543702:Mirn172 microRNA, Arabidopsis; D010759:Phosphorus Isotopes; D018749:RNA, Plant