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

Jia-Bao Long ,  Ying-Xin Liu ,  Qing-Feng Cao ,  Qiu-Ping Guo ,  Shu-Ya Yan ,  Xiang-Xian Meng

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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

    • 通讯作者: Xiang-Xian Meng,
  • 基金项目:

    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)

摘要: 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.

English

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  • 发布日期:  2015-05-31
  • 收稿日期:  2014-12-03
  • 网络出版日期:  2015-04-07
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