Rapid and sensitive detection of multiple microRNAs in cell lysate by low-fouling surface plasmon resonance biosensor
Language English Country Great Britain, England Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
25829219
DOI
10.1016/j.bios.2015.03.038
PII: S0956-5663(15)00190-6
Knihovny.cz E-resources
- Keywords
- DNA array, Erythrocyte lysate, Low-fouling surface chemistry, Polymer brushes, Surface plasmon resonance imaging, microRNA,
- MeSH
- Acrylamides chemistry MeSH
- Equipment Failure Analysis MeSH
- Coated Materials, Biocompatible chemical synthesis MeSH
- Biosensing Techniques instrumentation MeSH
- Equipment Design MeSH
- Cell Fractionation MeSH
- Complex Mixtures analysis MeSH
- MicroRNAs analysis chemistry genetics MeSH
- Polymers chemistry MeSH
- Surface Plasmon Resonance instrumentation MeSH
- Reproducibility of Results MeSH
- Sensitivity and Specificity MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Acrylamides MeSH
- Coated Materials, Biocompatible MeSH
- Complex Mixtures MeSH
- MicroRNAs MeSH
- poly(carboxybetaine acrylamide) MeSH Browser
- Polymers MeSH
We report an ultra-low fouling surface plasmon resonance imaging (SPRi) biosensor for the rapid simultaneous detection of multiple miRNAs in erythrocyte lysate (EL) at subpicomolar levels without need of RNA extraction. The SPRi chips were coated with ultra-low fouling functionalizable poly(carboxybetaine acrylamide) (pCBAA) brushes having optimized thicknesses and directly functionalized with amino-modified oligonucleotide probes. We have characterized the effect of the brush thickness on the probe loading capacity: a loading capacity of ~9.8×10(12) probes/cm(2) was achieved for pCBAA having a thickness of ~40 nm. The probe-functionalized sensor also exhibited a high resistance to fouling from ~90% EL samples (<2 ng/cm(2)). A two-step detection assay was employed for multiplexed miRNA detection in EL. Specifically, the assay consisted of (i) a sandwich-type hybridization of the probe-functionalized pCBAA with target miRNA in EL (bound to biotinylated oligonucleotides) and (ii) the capture of streptavidin-functionalized gold nanoparticles to the aforementioned biotinylated probes. We have demonstrated that this approach enables the detection of miRNAs in EL at concentrations as low as 0.5 pM. Finally, we have confirmed the detection of four endogenous miRNAs representing a set of potential miRNA biomarkers of myelodysplastic syndrome (MDS) in clinical EL samples (miR-16, miR-181, miR-34a, and miR-125b). The results revealed significantly higher levels of miR-16 in all the clinical EL samples compared to the other measured miRNAs.
References provided by Crossref.org
