利用汞离子(Hg2+)特异性功能核酸对Hg2+识别检测,其中模板主要包括与Hg2+特异性结合的识别区、形成G四链体的富G区以及由聚六乙二醇(Spacer18)链接的隔断区;靶序列主要包括5'端配对区和3'端富T区.模板与靶序列只有在Hg2+存在的条件下才能结合并引发延伸,进而使富G序列在模板上剥离下来,但由于Spacer18的隔断作用导致富G序列以单链形式存在,在特定环境下形成具有过氧化物模拟酶活性的G-四链体,催化H2 O2与2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二胺盐(ABTS)发生肉眼可见的颜色变化,进而对Hg2+进行定量测定.利用构建的Hg2+功能核酸生物传感器,35 min内可完成检测,线性范围为20~500 nmol/L,检出限达到16.5 nmol/L(3σ).实际水样中的加标回收率为98.5% ~103.5%.本方法操作简单、成本低、耗时短,在应急处理、实时环境检测等方面具有良好的应用价值.%Excessive mercury ions have negative effects on individual health. So, it is of great significance to develop a method for rapid, sensitive and specific detection of Hg2+. In this study, the method for detection of Hg2+ was developed based on specific T-Hg-T mismatche and G-quadruplex. The template sequence mainly included the recognition region which could combine with Hg2+ specifically, the rich G region which could form G-quadruplex, and the speacer 18 partition zone. The target sequence mainly included 5' ends combining area and 3' end T-rich region. Templates and targets could be combined and the elongation was triggered only in the presence of Hg2+. Furthermore, the G-rich sequences were stripped off the templates and could form a single-stranded structure, because Spacer 18 had partition function. The G-quadruplex was formed with the K+and hemin, and catalyzed H2 O2-2,2-azinobis (3-ethylbenzothiozoline)-6-sulfonic acid(ABTS) reaction with color variations. The detection could be done within 35 min, and the Hg2+ concentration exhibited a linear correlation with absorbance at 414 nm within range of 20-500 nmol/ L, with a detection limit of 16. 5 nmol/ L (3σ). The recoveries of Hg2+ spiked in tap water were 98. 5% - 103. 5% . The method exhibited the advantages such as simple operation, low cost and short time, and operation value in emergency treatment and real-time environmental detection.
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