Journal article
Sample Preparation-Free, Real-Time Detection of microRNA in Human Serum Using Piezoelectric Cantilever Biosensors at Attomole Level
Analytical chemistry (Washington), v 84(23), pp 10426-10436
04 Dec 2012
PMID: 23101954
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
A sensitive, selective, sample preparation-free method for near real-time detection of inicroRNA in buffer and human serum is given using gold (Au)-coated dynamic piezoelectric cantilever sensors. Sensor response to thiolated DNA probe chemisorption, hsa-let-7a hybridization, labeled-DNA hybridization, and Au nanoparticle-functionalized DNA hybridization was monitored continuously in flowing liquid samples using custom flow-cells. The assay showed successful detection of target let-7a with a dynamic range spanning 6 orders of magnitude (10 fM-1 nM) with a limit of detection of less than 10 attomoles (similar to 4 fM). The serum background had negligible effect on sensitivity relative to the results obtained in the buffer due to reduction in nonspecific binding caused by continuous sensor vibration. Both hybridization and nonspecific binding reduction were confirmed using fluorescence-based assays to support sensor-based results. The sensor-based method demonstrated excellent selectivity for the microRNA target in comparison with similar microRNA differing by only a single nucleotide (hsa-let-7c) and random microRNA sequences. Au nanoparticle-based amplification of sensor response was investigated and led to an order of magnitude improvement in the detection limit and a 128% amplification of sensor response over the entire dynamic range. Au nanopartide amplification was verified by scanning electron microscopy. The cantilever sensor-based microRNA assay provides competitive sensitivity with current microRNA detection methods and has the advantage of requiring no sample preparation, even when working with biological samples that contain a complex background.
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Details
- Title
- Sample Preparation-Free, Real-Time Detection of microRNA in Human Serum Using Piezoelectric Cantilever Biosensors at Attomole Level
- Creators
- Blake N. Johnson - Drexel UniversityRaj Mutharasan - Drexel University
- Publication Details
- Analytical chemistry (Washington), v 84(23), pp 10426-10436
- Publisher
- American Chemical Society; Washington, DC
- Number of pages
- 11
- Grant note
- CBET-0828987 / NSF; National Science Foundation (NSF)
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Chemical and Biological Engineering
- Web of Science ID
- WOS:000311815300040
- Scopus ID
- 2-s2.0-84870566312
- Other Identifier
- 991019170975004721
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InCites Highlights
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- Web of Science research areas
- Chemistry, Analytical