An isothermal system that couples ligand-dependent catalysis to ligand-independent exponential amplification.
An isothermal system that couples ligand-dependent catalysis to ligand-independent exponential amplification.
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DOI：
10.1021/ja111136d
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发表时间：
2011-03-09
影响因子：
15
通讯作者：
Joyce, Gerald F.
中科院分区：
文献类型：
--
作者：
Lam, Bianca J.;Joyce, Gerald F.
设计了一个系统，使定量，配体依赖指数扩增的各种配体，可以由RNA适配体识别。适体与一种催化两个寡核苷酸底物连接的RNA酶相连。这种反应的产物是另一种RNA酶，它在恒温下进行自我持续复制，拷贝数呈指数增长。配体的浓度决定了复制产物达到阈值浓度所需的时间。标准的阈值时间与配体浓度的关系图可用于确定未知样品中配体的浓度。该系统类似于定量PCR，将罕见的识别事件与随后的指数扩增联系起来，但与PCR不同的是，它可以应用于非核酸配体的定量检测。
A system was devised that enables quantitative, ligand-dependent exponential amplification for various ligands that can be recognized by an RNA aptamer. The aptamer is linked to an RNA enzyme that catalyzes the joining of two oligonucleotide substrates. The product of this reaction is another RNA enzyme that undergoes self-sustained replication at constant temperature, increasing in copy number exponentially. The concentration of the ligand determines the amount of time required for the replication products to reach a threshold concentration. A standardized plot of time to threshold versus ligand concentration can be used to determine the concentration of ligand in an unknown sample. This system is analogous to quantitative PCR, linking rare recognition events to subsequent exponential amplification, but unlike PCR can be applied to the quantitative detection of non-nucleic-acid ligands.
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