title: Surface plasmon resonance biosensor for rapid label-free detection of microribonucleic acid at subfemtomole level.
abstract: Microribonucleic acids (miRNAs) have been linked with various regulatory functions and disorders, such as cancers and heart diseases. They, therefore, present an important target for detection technologies for future medical diagnostics. We report here a novel method for rapid and sensitive miRNA detection and quantitation using surface plasmon resonance (SPR) sensor technology and a DNA*RNA antibody-based assay. The approach takes advantage of a novel high-performance portable SPR sensor instrument for spectroscopy of surface plasmons based on a special diffraction grating called a surface plasmon coupler and disperser (SPRCD). The surface of the grating is functionalized with thiolated DNA oligonucleotides which specifically capture miRNA from a liquid sample without amplification. Subsequently, an antibody that recognizes DNA*RNA hybrids is introduced to bind to the DNA*RNA complex and enhance sensor response to the captured miRNA. This approach allows detection of miRNA in less than 30 min at concentrations down to 2 pM with an absolute amount at high attomoles. The methodology is evaluated for analysis of miRNA from mouse liver tissues and is found to yield results which agree well with those provided by the quantitative polymerase chain reaction (qPCR).
authors.affiliation: Institute of Photonics and Electronics, Academy of Sciences of the Czech Republic, Chaberská 57, 18251 Prague, Czech Republic.
authors.forename: Hana
authors.initials: H
authors.lastname: Sípová
authors.forename: Shile
authors.initials: S
authors.lastname: Zhang
authors.forename: Aimée M
authors.initials: AM
authors.lastname: Dudley
authors.forename: David
authors.initials: D
authors.lastname: Galas
authors.forename: Kai
authors.initials: K
authors.lastname: Wang
authors.forename: Jirí
authors.initials: J
authors.lastname: Homola
mesh_terms: D000818:Animals; D000906:Antibodies; D015374:Biosensing Techniques / Q000379:methods*; D004247:DNA; D057230:Limit of Detection; D008099:Liver / Q000737:chemistry; D008722:Methods; D051379:Mice; D035683:MicroRNAs / Q000032:analysis*; D009693:Nucleic Acid Hybridization / Q000276:immunology; D012313:RNA; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D000906:Antibodies; D035683:MicroRNAs; D012313:RNA; D004247:DNA