A sensitive colorimetric assay system for nucleic acid detection based on isothermal signal amplification technology
- 8 July 2017
- journal article
- research article
- Published by Springer Science and Business Media LLC in Analytical and Bioanalytical Chemistry
- Vol. 409 (20), 4819-4825
- https://doi.org/10.1007/s00216-017-0425-4
Abstract
Rapid and accurate detection of microRNAs in biological systems is of great importance. Here, we report the development of a visual colorimetric assay which possesses the high amplification capabilities and high selectivity of the rolling circle amplification (RCA) method and the simplicity and convenience of gold nanoparticles used as a signal indicator. The designed padlock probe recognizes the target miRNA and is circularized, and then acts as the template to extend the target miRNA into a long single-stranded nucleotide chain of many tandem repeats of nucleotide sequences. Next, the RCA product is hybridized with oligonucleotides tagged onto gold nanoparticles. This interaction leads to the aggregation of gold nanoparticles, and the color of the system changes from wine red to dark blue according to the abundance of miRNA. A linear correlation between fluorescence and target oligonucleotide content was obtained in the range 0.3–300 pM, along with a detection limit of 0.13 pM (n = 7) and a RSD of 3.9% (30 pM, n = 9). The present approach provides a simple, rapid, and accurate visual colorimetric assay that allows sensitive biodetection and bioanalysis of DNA and RNA nucleotides of interest in biologically important samples. The colorimetric assay system for analyzing target oligonucleotidesKeywords
Funding Information
- the Excellent Young Teacher Program of China Medical University (YQ20160006)
- National Natural Science Foundation of China (81371896)
This publication has 30 references indexed in Scilit:
- Natural RNA circles function as efficient microRNA spongesNature, 2013
- Circular RNAs are a large class of animal RNAs with regulatory potencyNature, 2013
- Self-assembled RNA interference microsponges for efficient siRNA deliveryNature Materials, 2012
- miRNA Expression Profiling in Melanocytes and Melanoma Cell Lines Reveals miRNAs Associated with Formation and Progression of Malignant MelanomaJournal of Investigative Dermatology, 2009
- Stand-Alone Rolling Circle Amplification Combined with Capillary Electrophoresis for Specific Detection of Small RNAAnalytical Chemistry, 2009
- Rolling Circle Amplification: Applications in Nanotechnology and Biodetection with Functional Nucleic AcidsAngewandte Chemie-International Edition, 2008
- Integrin β3 expression is regulated by let-7a miRNA in malignant melanomaOncogene, 2008
- RAS Is Regulated by the let-7 MicroRNA FamilyCell, 2005
- Reduced Expression of the let-7 MicroRNAs in Human Lung Cancers in Association with Shortened Postoperative SurvivalCancer Research, 2004
- Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancersProceedings of the National Academy of Sciences of the United States of America, 2004