Orthopedics Department, The Third Affiliated Hospital of Chongqing Medical University (Gener Hospital), Chongqing, 401120, China.
Department of ICU, The Third Affiliated Hospital of Chongqing Medical University (Gener Hospital), Chongqing, 401120, China.
Anal Biochem. 2022 Mar 1;640:114543. doi: 10.1016/j.ab.2021.114543. Epub 2021 Dec 30.
Abnormal expression of microRNAs (miRNAs) is closely associated with a diverse of important biological processes, such as metastasis, myocardial ischemia and progression. Development of a facile and enzyme-free method for sensitive miRNA detection remains a huge challenge. Herein, we proposed a cross-catalytic circuit for trace miRNA detection by facilely integrating hairpin catalytic reaction (HCR) and DNAzyme biocatalyst through an ingenious feedback loop. The DNAzyme functional part was originally grafted in the designed hairpin structure probes, which would be released from hairpin structure probe and induce subsequent signal amplification after the recognition of target miRNA and the formation of double-strand DNA products. Through the dual signal amplification, the method exhibited a favorable detection sensitivity with a low of detection of 56 fM. These two indispensable catalytic reactions play vital roles in executing high-performance signal amplification, as demonstrated experimentally and theoretically.
异常表达的 microRNAs（miRNAs）与多种重要的生物学过程密切相关，如转移、心肌缺血和进展。开发一种简单且无酶的方法来灵敏地检测 miRNA 仍然是一个巨大的挑战。在此，我们通过巧妙的反馈环，将发夹催化反应（HCR）和 DNA 酶生物催化剂简单地整合在一起，提出了一种用于痕量 miRNA 检测的交叉催化电路。DNA 酶功能部分最初被嫁接在设计的发夹结构探针中，在目标 miRNA 的识别和双链 DNA 产物的形成后，它将从发夹结构探针中释放出来，并诱导随后的信号放大。通过双重信号放大，该方法表现出良好的检测灵敏度，检测下限低至 56 fM。这两种必不可少的催化反应在执行高性能信号放大方面发挥了重要作用，这在实验和理论上都得到了证明。
