Zou Mingyuan, Zhou Meiling, Ma Shuo, Zhang Chen, Xiao Feng, Wu Huina, GuliNaizhaer Abudushalamu, Yao Yuming, Chen Yaya, Cai Shijie, Fan Xiaobo, Zhao Fengfeng, Wu Guoqiu
Southeast University, Zhongda Hospital, Center of Clinical Laboratory Medicine, Medical School, Nanjing 210009, People's Republic of China.
Public Health School, Southeast University, Nanjing 210009, People's Republic of China.
iScience. 2023 Mar 13;26(4):106390. doi: 10.1016/j.isci.2023.106390. eCollection 2023 Apr 21.
Ever since the catalytic hairpin assembly (CHA) circuit has been highlighted as a powerful nucleic acid detection tool, nucleic acid detection methods based on CHA have been widely studied. However, the detection sensitivity of CHA-based methods is insufficient. The relatively high background signals resulting from the spontaneous reaction between the two hairpin probes is one of the major reasons for limiting the sensitivity of CHA. In this study, we established that the addition of deoxynucleotide triphosphates (dNTPs) to the reaction system can significantly reduce the background leakage of CHA. The dNTPs-CHA, coupled with a fluorescence lateral flow assay strip, is used for the rapid and highly sensitive detection of miRNA. It is capable of reliably detecting miRNA in serum samples down to a limit of 100 aM, which is an improvement in the lower detection limit by nearly five orders of magnitude compared to that of the pure CHA.
自从催化发夹组装（CHA）电路被视为一种强大的核酸检测工具以来，基于CHA的核酸检测方法得到了广泛研究。然而，基于CHA的方法检测灵敏度不足。两个发夹探针之间的自发反应产生的相对较高背景信号是限制CHA灵敏度的主要原因之一。在本研究中，我们发现向反应体系中添加三磷酸脱氧核苷酸（dNTPs）可显著降低CHA的背景泄漏。dNTPs-CHA与荧光侧向流动分析试纸条相结合，用于miRNA的快速、高灵敏检测。它能够可靠地检测血清样本中的miRNA，检测下限低至100 aM，与纯CHA相比，检测下限提高了近五个数量级。
