期刊
ACS NANO
卷 7, 期 5, 页码 3962-3969出版社
AMER CHEMICAL SOC
DOI: 10.1021/nn305789z
关键词
nanopore; single molecule; biosensor; nucleic acids; microRNA; miRNA; HIV-1 TAT; peptide; PNA; probe; cancer; diagnostics
类别
资金
- National Science Foundation [0546165]
- National Institutes of Health [GM079613]
- University of Missouri
- National Centre for Research Resources, National Institutes of Health [C06-RR-016489-01]
- Direct For Mathematical & Physical Scien
- Division Of Chemistry [0546165] Funding Source: National Science Foundation
- Div Of Molecular and Cellular Bioscience
- Direct For Biological Sciences [0920067] Funding Source: National Science Foundation
The nanopore sensor can detect cancer-derived nucleic acid biomarkers such as microRNAs (miRNAs), providing a noninvasive tool potentially useful in medical diagnostics. However, the nanopore-based detection of these biomarkers remains confounded by the presence of numerous other nucleic acid species found in biofluid extracts. Their nonspecific interactions with the nanopore inevitably contaminate the target signals, reducing the detection accuracy. Here we report a novel method that utilizes a polycationic peptide-PNA probe as the carrier for selective miRNA detection in the nucleic acid mixture. The cationic probe hybridized with microRNA forms a dipole complex, which can be captured by the pore using a voltage polarity that is opposite the polarity used to capture negatively charged nucleic acids. As a result, nontarget species are driven away from the pore opening, and the target miRNA can be detected accurately without interference. In addition, we demonstrate that the PNA probe enables accurate discrimination of miRNAs with single-nucleotide difference. This highly sensitive and selective nanodielectrophoresis approach can be applied to the detection of clinically relevant nucleic acid fragments in complex samples.
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