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.

中文翻译：

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高选择性和高灵敏度的连接依赖滚环扩增法测定 ATP

迫切需要开发可靠检测ATP的方法，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，目前可用于 ATP 传感的方法强烈依赖于使用昂贵和复杂的仪器或使用具有中等灵敏度和选择性的 ATP 适体。为了避免这些缺点，我们在此提出了一种有效的 ATP 检测方法，通过将高度特异性的 ATP 依赖性连接反应与由滚环扩增 (RCA) 执行的双阶段信号放大技术相结合，以及随后制备的 DNAzymes 准备用于催化裂解和荧光从分子信标 (MB) 生成信号。检测限低至 35 pM，线性范围为 0.05 nM 至 200 nM。更重要的是，传感策略可以有效地将 ATP 与其类似物区分开来，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、免洗和等温方便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 和其他生物分子铺平了新的途径。