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A cascade signal amplification strategy for sensitive and label-free DNA detection based on Exo III-catalyzed recycling coupled with rolling circle amplification.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Liu, Xingti Xue, Qingwang Ding, Yongshun Zhu, Jing Wang, Lei Jiang, Wei |
| Copyright Year | 2014 |
| Abstract | A sensitive and label-free fluorescence assay for DNA detection has been developed based on cascade signal amplification combining exonuclease III (Exo III)-catalyzed recycling with rolling circle amplification. In this assay, probe DNA hybridized with template DNA was coupled onto magnetic nanoparticles to prepare a magnetic bead-probe (MNB-probe)-template complex. The complex could hybridize with the target DNA, which transformed the protruding 3' terminus of template DNA into a blunt end. Exo III could then digest template DNA, liberating the MNB-probe and target DNA. The intact target DNA then hybridized with other templates and released more MNB-probes. The liberated MNB-probe captured the primer, circular DNA and then initiated the rolling circle amplification (RCA) reaction, realizing a cascade signal amplification. Using this cascade amplification strategy, a sensitive DNA detection method was developed which was superior to many existing Exo III-based signal amplification methods. Moreover, N-methyl mesoporphyrin IX, which had a pronounced structural selectivity for the G-quadruplex, was used to combine with the G-quadruplex RCA products and generate a fluorescence signal, avoiding the need for any fluorophore-label probes. The spike and recovery experiments in a human serum sample indicated that our assay also had great potential for DNA detection in real biological samples. |
| File Format | PDF HTM / HTML |
| DOI | 10.1039/c4an00389f |
| Alternate Webpage(s) | http://pubs.rsc.org/en/content/getauthorversionpdf/C4AN00389F |
| PubMed reference number | 24752174 |
| Alternate Webpage(s) | https://doi.org/10.1039/c4an00389f |
| Journal | Medline |
| Volume Number | 139 |
| Issue Number | 11 |
| Journal | The Analyst |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
