Detection of bacterial 16S rRNA using a molecular beacon-based X sensor

    Authors

    Y. V. Gerasimova;D. M. Kolpashchikov

    Comments

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    Abbreviated Journal Title

    Biosens. Bioelectron.

    Keywords

    Hybridization probes; Nucleic acid detection; Molecular beacon probe; 16S rRNA; DNA crossover; PEPTIDE NUCLEIC-ACIDS; OLIGONUCLEOTIDE PROBES; IN-SITU; LIVING CELLS; SNP ANALYSIS; HYBRIDIZATION; MICROARRAYS; ACCESSIBILITY; DIAGNOSTICS; CAPTURE; Biophysics; Biotechnology & Applied Microbiology; Chemistry, Analytical; Electrochemistry; Nanoscience & Nanotechnology

    Abstract

    We demonstrate how a long structurally constrained RNA can be analyzed in homogeneous solution at ambient temperatures with high specificity using a sophisticated biosensor. The sensor consists of a molecular beacon probe as a signal reporter and two DNA adaptor strands, which have fragments complementary to the reporter and to the analyzed RNA. One adaptor strand uses its long RNA-binding arm to unwind the RNA secondary structure. Second adaptor strand with a short RNA-binding arm hybridizes only to a completely complementary site, thus providing high recognition specificity. Overall the three-component sensor and the target RNA form a four-stranded DNA crossover (X) structure. Using this sensor, Escherichia coli16 S rRNA was detected in real time with the detection limit of similar to 0.17 nM. The high specificity of the analysis was proven by differentiating Bacillus subtilis from E. coli 16 S rRNA sequences. The sensor responds to the presence of the analyte within seconds. (C) 2012 Elsevier B.V. All rights reserved.

    Journal Title

    Biosensors & Bioelectronics

    Volume

    41

    Publication Date

    1-1-2013

    Document Type

    Article

    Language

    English

    First Page

    386

    Last Page

    390

    WOS Identifier

    WOS:000314191300056

    ISSN

    0956-5663

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