Proximity-enabled Bidirectional Enzymatic Repairing Amplification for Ultrasensitive Fluorescence Sensing of Adenosine Triphosphate,Analytica Chimica Acta

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Proximity-enabled Bidirectional Enzymatic Repairing Amplification for Ultrasensitive Fluorescence Sensing of Adenosine Triphosphate
Analytica Chimica Acta ( IF 5.7 ) Pub Date : 2020-04-01 , DOI: 10.1016/j.aca.2020.01.006
Shasha Li ₁ , Su Liu ₂ , Jingfeng Wang ₁ , Yihan Zhao ₁ , Rufeng Zhang ₂ , Xiaonan Qu ₂ , Yu Wang ₁ , Jiadong Huang ₃ , Jinghua Yu ₄

Affiliation

A novel fluorescence sensing strategy for ultrasensitive and highly specific detection of adenosine triphosphate (ATP) has been developed by the combination of the proximity ligation assay with bidirectional enzymatic repairing amplification (BERA). The strategy relies on proximity binding-triggered the release of palindromic tail that initiates bidirectional cyclic enzymatic repairing amplification reaction with the aid of polymerase and two DNA repairing enzymes, uracil-DNA glycosylase (UDG) and endonuclease IV (Endo IV). A fluorescence-quenched hairpin probe with a palindromic tail at the 3' end is skillfully designed that functions as not only the recognition element, primer, and polymerization template for BERA but also the indicator for fluorescence signal output. On the basis of the amplification strategy, this biosensor displays excellent sensitivity and selectivity for ATP detection with an outstanding detection limit of 0.81 pM. Through simultaneously enhancing the target response signal value and reducing nonspecific background, this work deducted the background effect, and showed high sensitivity and reproducibility. Moreover, our biosensor also shows promising potential in real sample analysis. Therefore, the proximity-enabled BERA strategy indeed creates a simple and valuable fluorescence sensing platform for ATP identification and related disease diagnosis and biomedical research.

中文翻译：

用于三磷酸腺苷超灵敏荧光传感的近程双向酶修复放大

通过将邻近连接测定与双向酶促修复扩增 (BERA) 相结合，开发了一种用于超灵敏和高特异性检测三磷酸腺苷 (ATP) 的新型荧光传感策略。该策略依赖于邻近结合触发回文尾的释放，在聚合酶和两种 DNA 修复酶尿嘧啶-DNA 糖基化酶 (UDG) 和核酸内切酶 IV (Endo IV) 的帮助下启动双向循环酶修复扩增反应。巧妙地设计了3'端带有回文尾的荧光猝灭发夹探针，它既是BERA的识别元件、引物和聚合模板，又是荧光信号输出的指示剂。在放大策略的基础上，该生物传感器对 ATP 检测具有出色的灵敏度和选择性，检测限为 0.81 pM。该工作通过同时提高目标响应信号值和降低非特异性背景，扣除背景效应，表现出较高的灵敏度和重现性。此外，我们的生物传感器在实际样品分析中也显示出巨大的潜力。因此，支持接近度的 BERA 策略确实为 ATP 识别和相关疾病诊断和生物医学研究创建了一个简单而有价值的荧光传感平台。并显示出高灵敏度和重现性。此外，我们的生物传感器在实际样品分析中也显示出巨大的潜力。因此，接近启用的BERA策略确实为ATP识别和相关疾病诊断和生物医学研究创建了一个简单而有价值的荧光传感平台。并显示出高灵敏度和重现性。此外，我们的生物传感器在实际样品分析中也显示出有希望的潜力。因此，支持接近度的 BERA 策略确实为 ATP 识别和相关疾病诊断和生物医学研究创建了一个简单而有价值的荧光传感平台。

更新日期：2020-04-01

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