滚动圆复制
化学
核酸
DNA
生物系统
环介导等温扩增
指数函数
灵敏度(控制系统)
指数增长
多重位移放大
脱氧核酶
底漆(化妆品)
寡核苷酸
生物物理学
信号(编程语言)
计算生物学
A-DNA
微流控
串联
检出限
聚合酶
DNA聚合酶
动力学
聚合酶链反应
劈理(地质)
多路复用
纳米技术
作者
Jinhua Shang,Mengdi Yu,Yifei Wang,Shanshan Yu,XianZheng ZHANG,Fuan Wang,Yingfu Li,Jinhua Shang,Mengdi Yu,Yifei Wang,Shanshan Yu,XianZheng ZHANG,Fuan Wang,Yingfu Li
摘要
Rolling circle amplification (RCA) is a powerful isothermal strategy for nucleic acid detection, but its linear kinetics and dependence on externally supplied primers limit its sensitivity and programmability. Here, we report an exponential RCA (E-RCA) platform that integrates primer regeneration and signal amplification into a single DNA-encoded system. The design uses a circular DNA template encoding the I-R3 self-cleaving DNAzyme sequence; during RCA, tandem I-R3 units are generated within the DNA amplicons, which then catalyze site-specific cleavage to release new primers. This self-sustained (RCA ↔ DNAzyme) amplification circuit enables robust exponential signal growth using only a circular DNA probe and a DNA polymerase without requiring external primers or protein enzymes. We elucidate the mechanism through biochemical experiments and kinetic modeling and validate the system in multiplexed intracellular microRNA imaging and quantitative dual-marker profiling of clinical breast cancer tissues. The E-RCA strategy achieved high diagnostic accuracy (AUC = 0.914; specificity = 100%; sensitivity = 81.3%), demonstrating its potential for sensitive, programmable, and autonomous molecular analysis in both biological and clinical contexts.
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