Citation: Jia-Bao Long, Ying-Xin Liu, Qing-Feng Cao, Qiu-Ping Guo, Shu-Ya Yan, Xiang-Xian Meng. Sensitive and enzyme-free detection for single nucleotide polymorphism using microbead-assisted toehold-mediated strand displacement reaction[J]. Chinese Chemical Letters, ;2015, 26(8): 1031-1035. doi: 10.1016/j.cclet.2015.05.036 shu

Sensitive and enzyme-free detection for single nucleotide polymorphism using microbead-assisted toehold-mediated strand displacement reaction

1.
a College of Biology, State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082, China;
2.
b Department of Bioengineering and Environment Science, Changsha University, Changsha 410003, China

Corresponding author: Xiang-Xian Meng,
Received Date: 3 December 2014
Available Online: 7 April 2015
Fund Project: This work is supported by National Natural Science Foundation of China (No. 21275043) (No. 21275043)National Basic Research Program of China under Grants (No. 2009CB421601). (No. 2009CB421601)

This report described a free-enzyme, convenient and inexpensive genotyping biosensor capable of detecting single nucleotide polymorphism at normal temperature based on the combination of toeholdmediated strand displacement reaction (toehold-SDR) and microbead-capture technique. The biosensor consists of a pre-hybridized strand formed by a reporter probe and a capture probe. In the presence of a mutant sequence, there is no toehold-mediated strand displacement and the reporter probe cannot be released from the pre-hybridized strand. Microbeads capture the fluorescent pre-hybridized strand through biotin-streptavidin interaction, so microbeads give out significant fluorescence signal, while there is no fluorescence in the solution. However, in the presence of a matched target, the strand displacement is effectively initiated and the reporter probe is released from pre-hybridized strand. After addingmicrobeads, the solution produces bright fluorescence, while microbeads have no obvious signal. Genotypes are identified conveniently according to the fluorescence intensity of the solution. The method provides a simple and inexpensive strategy to detect point mutation. Moreover, this biosensor shows the linear relationship in the range of 1-40 nmol/L and reaches a detection limit of 0.3 nmol/L.
- Enzyme-free,
- Microbead-assisted,
- Toehold-mediated strand displacement,
- Point mutation
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沈阳化工大学材料科学与工程学院沈阳 110142