| DOI | Resolve DOI: https://doi.org/10.1021/ac2016213 |
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| Author | Search for: Khan, N.; Search for: Cheng, J.1; Search for: Pezacki, J.P.1; Search for: Berezovski, M.V. |
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| Affiliation | - National Research Council of Canada. NRC Steacie Institute for Molecular Sciences
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| Format | Text, Article |
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| Subject | Affinity capillary electrophoresis; Blood samples; Blood serum; Cancer classification; Detection limits; Double-stranded RNA; Magnetic beads; MicroRNAs; Potential sources; Pre-concentration; Selective binding; Single-stranded DNA; Single-stranded dna binding proteins; Blood; Capillary electrophoresis; Diseases; DNA; Polymerase chain reaction; Probes; Proteins; RNA; Tissue; Blood vessels; DNA binding protein; double stranded RNA; fluorescent dye; microRNA; MIRN122 microRNA, human; RNA binding protein; single stranded DNA; biosynthesis; blood; capillary electrophoresis; chemical analysis; chemistry; limit of detection; metabolism; methodology; nucleic acid hybridization; RNA probe; Chemistry Techniques, Analytical; DNA, Single-Stranded; DNA-Binding Proteins; Electrophoresis, Capillary; Fluorescent Dyes; Limit of Detection; MicroRNAs; Nucleic Acid Hybridization; RNA Probes; RNA, Double-Stranded; RNA-Binding Proteins |
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| Abstract | MicroRNAs (miRNAs) are small (∼22 nt) regulatory RNAs that are frequently deregulated in cancer and have shown promise as tissue- and blood-based biomarkers for cancer classification and prognostication. Here we present a protein-facilitated affinity capillary electrophoresis (ProFACE) assay for rapid quantification of miRNA levels in blood serum using single-stranded DNA binding protein (SSB) and double-stranded RNA binding protein (p19) as separation enhancers. The method utilizes either the selective binding of SSB to a single-stranded DNA/RNA probe or the binding of p19 to miRNA-RNA probe duplex. For the detection of ultralow amounts of miRNA without polymerase chain reaction (PCR) amplification in blood samples we apply off-line preconcentration of synthetic miRNA-122 from serum by p19-coated magnetic beads followed by online sample stacking in the ProFACE assay. The detection limit is 0.5 fM or 30 000 miRNA molecules in 1 mL of serum as a potential source of naïve miRNAs. © 2011 American Chemical Society. |
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| Publication date | 2011 |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| NPARC number | 21271260 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | 26b53f0f-0ba1-451b-b358-3ccbc118b3fd |
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| Record created | 2014-03-24 |
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| Record modified | 2020-04-21 |
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