Real-Time Detection of Isothermal Amplification Reactions with Thermostable Catalytic Hairpin Assembly,Journal of the American Chemical Society

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Real-Time Detection of Isothermal Amplification Reactions with Thermostable Catalytic Hairpin Assembly
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2013-05-09 , DOI: 10.1021/ja4023978
Yu Sherry Jiang ₁ , Bingling Li , John N Milligan , Sanchita Bhadra , Andrew D Ellington

Affiliation

Catalytic hairpin assembly (CHA) is an enzyme-free amplification method that has previously proven useful in amplifying and transducing signals at the terminus of nucleic acid amplification reactions. Here, for the first time, we engineered CHA to be thermostable from 37 to 60 °C and in consequence have generalized its application to the real-time detection of isothermal amplification reactions. CHA circuits were designed and optimized for both high- and low-temperature rolling circle amplification (RCA) and strand displacement amplification (SDA). The resulting circuits not only increased the specificity of detection but also improved the sensitivity by as much as 25- to 10000-fold over comparable real-time detection methods. These methods have been condensed into a set of general rules for the design of thermostable CHA circuits with high signals and low noise.

中文翻译：

使用热稳定催化发夹组件实时检测等温扩增反应

催化发夹组装 (CHA) 是一种无酶扩增方法，此前已证明该方法可用于在核酸扩增反应的终点放大和转导信号。在这里，我们首次将 CHA 设计为在 37 至 60 °C 之间具有热稳定性，因此将其应用推广到等温扩增反应的实时检测。CHA 电路针对高温和低温滚环扩增 (RCA) 和链置换扩增 (SDA) 进行了设计和优化。由此产生的电路不仅提高了检测的特异性，而且与可比的实时检测方法相比，灵敏度提高了 25 到 10000 倍。

更新日期：2013-05-09

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