NHC Key Laboratory of Carcinogenesis and Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Cancer Research Institute and School of Basic Medical Science, Central South University, Changsha 410078, P. R. China.
Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Disease Genome Research Center, the Third Xiangya Hospital, Central South University, Changsha 410013, P. R. China.
Analyst. 2021 Oct 25;146(21):6605-6614. doi: 10.1039/d1an01115d.
It is highly demanded to develop methods for the reliable detection of ATP, which plays an extremely important role in clinical diagnosis, biomedical engineering, and food chemistry. However, the methods currently available for ATP sensing strongly rely on the utilization of expensive and sophisticated instruments or the use of ATP aptamers with mediocre sensitivity and selectivity. To circumvent these drawbacks, we herein propose an efficient method for ATP detection by integrating highly specific ATP-dependent ligation reaction with dual-stage signal amplification techniques executed by rolling circle amplification (RCA) and the subsequently fabricated DNAzymes ready for the catalytic cleavage and fluorescence signal generation from molecular beacons (MBs). The detection limit is down to 35 pM with a linear range from 0.05 nM to 200 nM. More importantly, the sensing strategy can effectively discriminate ATP from its analogues and the results from the spiked human serum albumin (HSA) samples further confirm the reliability for practical applications. Considering the high sensitivity and selectivity, wash-free and isothermal convenience, and the simplicity in probe design, the strategy reported herein paves a new avenue for the effective determination of ATP and other biomolecules in fundamental and applied research.
开发可靠的 ATP 检测方法的需求非常高，ATP 在临床诊断、生物医学工程和食品化学中起着极其重要的作用。然而，目前用于 ATP 传感的方法强烈依赖于使用昂贵且复杂的仪器，或使用具有中等灵敏度和选择性的 ATP 适体。为了规避这些缺点，我们在此提出了一种通过整合高度特异性的 ATP 依赖性连接反应与双阶段信号放大技术来检测 ATP 的有效方法，该方法通过滚环扩增（RCA）和随后制备的 DNA 酶进行执行，用于从分子信标（MB）进行催化切割和荧光信号产生。检测限低至 35 pM，线性范围为 0.05 nM 至 200 nM。更重要的是，该传感策略可以有效地将 ATP 与其类似物区分开来，并且来自加标人血清白蛋白（HSA）样品的结果进一步证实了实际应用的可靠性。考虑到高灵敏度和选择性、免洗和等温便利性以及探针设计的简单性，本文报道的策略为在基础和应用研究中有效测定 ATP 和其他生物分子开辟了新途径。
