Establishment of a universal and rational gene detection strategy through three-way junction-based remote transduction.
Establishment of a universal and rational gene detection strategy through three-way junction-based remote transduction.
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DOI：
10.1039/c7sc03190d
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发表时间：
2018-01-21
期刊：
影响因子：
8.4
通讯作者：
Li B
中科院分区：
文献类型：
--
作者：
Tang Y;Lu B;Zhu Z;Li B
聚合酶链式反应和多次等温扩增能够实现超基因扩增。聚合酶链式反应和多次等温扩增能够实现超基因扩增。遗憾的是，大多数常用的转导方法，如染料染色和Taqman类探针，仍然存在着信号错误或探针设计困难等缺点，或者与多项分析不兼容。在这里，通过基于三向连接的远程转导，将等温扩增转化为无酶的链置换电路，建立了一种通用而合理的基因检测策略。引入辅助转导探针，与靶单链核酸形成部分杂交。经过系统优化后，混合体可以作为关联触发器，通过跨越三向连接的链置换反应来激活下游电路检测器。这样，通过放大-换能器识别和放大-回路反应，检测的选择性可以得到双重保证。根据目标，通过仅在一个组件上简单地移动10-12个nT，良好优化的电路可以立即应用于新的目标检测。更重要的是，这一特性首次实现了单一反应中的多分析和逻辑分析，共享一个荧光报告。在一个可应用的模型中，微量的克隆氏菌和肠杆菌基因已经与没有细菌或只有一种细菌的样品明显地区分开来，具有超高的灵敏度和选择性。
The polymerase chain reaction and many isothermal amplifications are able to achieve super gene amplification. The polymerase chain reaction and many isothermal amplifications are able to achieve super gene amplification. Unfortunately, most commonly-used transduction methods, such as dye staining and Taqman-like probing, still suffer from shortcomings including false signals or difficult probe design, or are incompatible with multi-analysis. Here a universal and rational gene detection strategy has been established by translating isothermal amplicons to enzyme-free strand displacement circuits via three-way junction-based remote transduction. An assistant transduction probe was imported to form a partial hybrid with the target single-stranded nucleic acid. After systematic optimization the hybrid could serve as an associative trigger to activate a downstream circuit detector via a strand displacement reaction across the three-way junction. By doing so, the detection selectivity can be double-guaranteed through both amplicon–transducer recognition and the amplicon–circuit reaction. A well-optimized circuit can be immediately applied to a new target detection through simply displacing only 10–12 nt on only one component, according to the target. More importantly, this property for the first time enables multi-analysis and logic-analysis in a single reaction, sharing a single fluorescence reporter. In an applicable model, trace amounts of Cronobacter and Enterobacteria genes have been clearly distinguished from samples with no bacteria or one bacterium, with ultra-high sensitivity and selectivity.
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影响因子：
4.6
作者：
Du Y;Hughes RA;Bhadra S;Jiang YS;Ellington AD;Li B
通讯作者：
Li B
无酶DNA电路对多种检测方法的有理模块化适应。
影响因子：
14.9
作者：
Li B;Ellington AD;Chen X
通讯作者：
Chen X
DOI：
10.1073/pnas.0407024101
发表时间：
2004-10-26
影响因子：
11.1
作者：
Dirks, RM;Pierce, NA
通讯作者：
Pierce, NA
影响因子：
62.1
作者：
Liu, Juewen;Cao, Zehui;Lu, Yi
通讯作者：
Lu, Yi
影响因子：
14.9
作者：
Ang YS;Tong R;Yung LY
通讯作者：
Yung LY