Rapid and sensitive detection of multiple microRNAs in cell lysate by low-fouling surface plasmon resonance biosensor
Jazyk angličtina Země Velká Británie, Anglie Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
25829219
DOI
10.1016/j.bios.2015.03.038
PII: S0956-5663(15)00190-6
Knihovny.cz E-zdroje
- Klíčová slova
- DNA array, Erythrocyte lysate, Low-fouling surface chemistry, Polymer brushes, Surface plasmon resonance imaging, microRNA,
- MeSH
- akrylamidy chemie MeSH
- analýza selhání vybavení MeSH
- biokompatibilní potahované materiály chemická syntéza MeSH
- biosenzitivní techniky přístrojové vybavení MeSH
- design vybavení MeSH
- frakcionace buněk MeSH
- komplexní směsi analýza MeSH
- mikro RNA analýza chemie genetika MeSH
- polymery chemie MeSH
- povrchová plasmonová rezonance přístrojové vybavení MeSH
- reprodukovatelnost výsledků MeSH
- senzitivita a specificita MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- akrylamidy MeSH
- biokompatibilní potahované materiály MeSH
- komplexní směsi MeSH
- mikro RNA MeSH
- poly(carboxybetaine acrylamide) MeSH Prohlížeč
- polymery 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.
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