脱氧核酶
次氯酸
核酸
化学
检出限
劈理(地质)
荧光
氧化磷酸化
生物物理学
生物化学
组合化学
色谱法
材料科学
生物
物理
量子力学
断裂(地质)
复合材料
作者
Jiaqi Xu,Xi Zhou,Hui He,Shanni Li,Changbei Ma
标识
DOI:10.1016/j.saa.2023.123243
摘要
Hypochlorous acid (HClO) is a crucial active oxygen component and one of the innate immunity's barrier substances in the body. Abnormal fluctuations in HClO concentration can lead to increased oxidative stress, cellular dysfunction, and the onset of various diseases. Thus, developing convenient, affordable, efficient, and sensitive methods to monitor HClO concentration in healthcare and pathophysiology research is highly significant. In this study, we developed a novel fluorescence strategy for HClO detection based on nucleic acid oxidative cleavage and Pb2+-dependent DNAzyme. By introducing a phosphorothioate site in the hairpin-structured nucleic acid sequence, the nucleic acid probe specifically recognized HClO and underwent oxidative cleavage. Upon cleavage, the enzyme strand is liberated, forming DNAzyme. This DNAzyme then cleaves the substrate strand, liberating the initially quenched fluorescent dyes and generating a turn-on fluorescent signal. The enzyme strand produced by the oxidative cleavage of HClO can be repeatedly utilized, thus realizing the cyclic signal amplification to reduce background noise. We verified the detection mechanism of this strategy through stepwise fluorescence spectroscopy analysis and electrophoresis. Under optimal experimental conditions, the method achieved a detection limit of 5.38 nM and a linear range of 1 nM-800 nM. This method demonstrated exceptional performance in actual biological sample testing and presented an exciting opportunity for expanded utilization in clinical diagnosis and medical research.
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