State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
Biosens Bioelectron. 2013 Sep 15;47:324-8. doi: 10.1016/j.bios.2013.03.020. Epub 2013 Mar 29.
We developed a novel electrochemical strategy for ultrasensitive DNA detection using a dual amplification strategy based on the circular strand-displacement polymerase reaction (CSDPR) and the hybridization chain reaction (HCR). In this assay, hybridization of hairpin-shaped capture DNA to target DNA resulted in a conformational change of the capture DNA with a concomitant exposure of its stem. The primer was then hybridized with the exposed stem and triggered a polymerization reaction, allowing a cyclic reaction comprising release of target DNA, hybridization of target with remaining capture DNA, polymerization initiated by the primer. Furthermore, the free part of the primer propagated a chain reaction of hybridization events between two DNA hairpin probes with biotin labels, enabling an electrochemical reading using the streptavidin-alkaline phosphatase. The proposed biosensor showed to have very high sensitivity and selectivity with a dynamic response range through 10fM to 1nM, and the detect limit was as low as 8fM. The proposed strategy could have the potential for molecular diagnostics in complex biological systems.
我们开发了一种新的电化学策略,用于使用基于循环链置换聚合反应(CSDPR)和杂交链式反应(HCR)的双重扩增策略进行超灵敏 DNA 检测。在该测定中,发夹状捕获 DNA 与靶 DNA 的杂交导致捕获 DNA 的构象变化,同时暴露其茎。然后,引物与暴露的茎杂交并引发聚合反应,允许包括释放靶 DNA、靶与剩余捕获 DNA 杂交、由引物引发的聚合反应的循环反应。此外,引物的游离部分在具有生物素标记的两个 DNA 发夹探针之间的杂交事件中引发链反应,从而可以使用链霉亲和素-碱性磷酸酶进行电化学生物传感器。该生物传感器具有非常高的灵敏度和选择性,动态响应范围从 10fM 到 1nM,检测限低至 8fM。该策略有可能用于复杂生物系统中的分子诊断。