期刊
ANALYTICAL BIOCHEMISTRY
卷 412, 期 2, 页码 165-172出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2011.01.030
关键词
MicroRNAs; Surface plasmon resonance; Quantification; Enzyme immunoassay; p19
资金
- Genomics and Health Initiative of the National Research Council of Canada
- Natural Sciences and Engineering Research Council of Canada (NSERC)
- CIHR
MicroRNAs (miRNAs) are endogenous posttranscriptional regulators found in all metazoa and play crucial roles in virtually all cellular processes. Their aberrant expression has been linked to several diseased states; therefore, techniques capable of sensitive and specific profiling of the miRNA milieu will have significant application in prognostics, diagnostics, and therapeutics. Here we present a method for rapid quantification of miRNA levels using p19, a tombusvirus-encoded suppressor of RNA interference with sequence-independent and size-selective affinity toward 19-bp RNA duplexes. We present a surface plasmon resonance (SPR)-based miRNA sensing method where RNA probes are immobilized on gold surfaces demonstrating p19's utility in recognition of miRNA-bound probes. This allows detection of miRNAs in the low nanomolar range. To increase the sensitivity, a bead-based enzyme immunoassay was performed, and this technique displays a lower detection limit of 1 fmol and a linear dynamic range from 1 pmol to 1 fmol. Crown Copyright (C) 2011 Published by Elsevier Inc. All rights reserved.
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