Sensitive fluorescence detection of nucleic acids based on isothermal circular strand-displacement polymerization reaction
Sensitive fluorescence detection of nucleic acids based on isothermal circular strand-displacement polymerization reaction
复制标题
基于等温环链置换聚合反应的核酸灵敏荧光检测
DOI：
10.1093/nar/gkn1024
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发表时间：
2009-02-01
影响因子：
14.9
通讯作者：
Li, Huimin
中科院分区：
文献类型：
--
作者：
Guo, Qiuping;Yang, Xiaohai;Li, Huimin
我们开发了一种基于等温链置换聚合反应的灵敏 DNA 扩增检测方法。该方法同时利用了DNA的杂交特性和聚合酶的链置换特性。重要的是，我们证明我们的方法会产生由目标激活的循环聚合反应，这基本上允许它进行自我检测。从功能上讲，该 DNA 系统由发夹荧光探针、短引物和聚合酶组成。识别并与目标 ssDNA 杂交后，发夹探针的茎打开，然后打开的探针与引物退火并引发聚合反应。在此聚合反应过程中，合成了互补 DNA，并置换了杂交靶标。最后，移位的目标识别并与另一个探针杂交，触发下一轮聚合反应，达到6.4 × 10−15 M的目标检测限。
Here we have developed a sensitive DNA amplified detection method based on isothermal strand-displacement polymerization reaction. This method takes advantage of both the hybridization property of DNA and the strand-displacement property of polymerase. Importantly, we demonstrate that our method produces a circular polymerization reaction activated by the target, which essentially allows it to self-detect. Functionally, this DNA system consists of a hairpin fluorescence probe, a short primer and polymerase. Upon recognition and hybridization with the target ssDNA, the stem of the hairpin probe is opened, after which the opened probe anneals with the primer and triggers the polymerization reaction. During this process of the polymerization reaction, a complementary DNA is synthesized and the hybridized target is displaced. Finally, the displaced target recognizes and hybridizes with another probe, triggering the next round of polymerization reaction, reaching a target detection limit of 6.4 × 10−15 M.