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Amplification of the Fluorescence Signal with Clustered Regularly Interspaced Short Palindromic Repeats-Cas12a Based on Au Nanoparticle-DNAzyme Probe and On-Site Detection of Pb2+ Via the Photonic Crystal Chip
ACS Sensors ( IF 8.2 ) Pub Date : 2022-04-28 , DOI: 10.1021/acssensors.2c00516 Yu-Yao Li 1 , Hao-Dong Li 2 , Wen-Kai Fang 1 , Da Liu 1 , Meng-Han Liu 1 , Ming-Qiu Zheng 1 , Li-Ling Zhang 1 , He Yu 1 , Hong-Wu Tang 1
ACS Sensors ( IF 8.2 ) Pub Date : 2022-04-28 , DOI: 10.1021/acssensors.2c00516 Yu-Yao Li 1 , Hao-Dong Li 2 , Wen-Kai Fang 1 , Da Liu 1 , Meng-Han Liu 1 , Ming-Qiu Zheng 1 , Li-Ling Zhang 1 , He Yu 1 , Hong-Wu Tang 1
Affiliation
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Although great headway has been made in DNAzyme-based detection of Pb2+, its adaptability, sensitivity, and accessibility in complex media still need to be improved. For this, we introduce new ways to surmount these hurdles. First, a spherical nucleic acid (SNA) fluorescence probe (Au nanoparticles-DNAzyme probe) is utilized to specifically identify Pb2+ and its suitability for precise detection of Pb2+ in complex samples due to its excellent nuclease resistance. Second, the sensitivity of Pb2+ detection is greatly enhanced via the use of a clustered regularly interspaced short palindromic repeats-Cas12a with target recognition accuracy to amplify the fluorescent signal upon the trans cleavage of the SNA (signal probe), and the limit of detection reaches as low as 86 fM. Third, we boost the fluorescence on photonic crystal chips with a bionic periodic arrangement by employing a straightforward detection device (smartphone and portable UV lamp) to achieve on-site detection of Pb2+ with the limit of detection as low as 24 pM. Based on the abovementioned efforts, the modified Pb2+ fluorescence sensor has the advantages of higher sensitivity, better specificity, accessibility, less sample consumption, and so forth. Moreover, it can be applied to accurately detect Pb2+ in complex biological or environmental samples, which is of great promise for widespread applications.
中文翻译：
基于金纳米粒子-脱氧核糖核酸酶探针的成簇规则间隔短回文重复序列-Cas12a的荧光信号放大和通过光子晶体芯片现场检测Pb2+
尽管基于脱氧核酶的Pb 2+检测取得了很大进展，但其在复杂介质中的适应性、灵敏度和可及性仍有待提高。为此，我们引入了克服这些障碍的新方法。首先，利用球形核酸（SNA）荧光探针（Au nanoparticle-DNAzyme probe）对Pb 2+进行特异性鉴定，并因其优异的抗核酸酶性而适用于复杂样品中Pb 2+的精确检测。二、Pb 2+的灵敏度通过使用具有目标识别精度的成簇的规则间隔短回文重复序列-Cas12a 来放大 SNA（信号探针）反式切割时的荧光信号，大大增强了检测能力，检测限低至 86 fM。第三，我们通过使用简单的检测设备（智能手机和便携式紫外灯）来增强具有仿生周期性排列的光子晶体芯片上的荧光，以实现 Pb 2+的现场检测，检测限低至 24 pM。基于上述努力，改进后的Pb 2+荧光传感器具有更高的灵敏度、更好的特异性、可及性、更少的样品消耗等优点。此外，它还可用于准确检测 Pb2+在复杂的生物或环境样品中，这对于广泛的应用具有很大的前景。
更新日期：2022-04-28
中文翻译：
基于金纳米粒子-脱氧核糖核酸酶探针的成簇规则间隔短回文重复序列-Cas12a的荧光信号放大和通过光子晶体芯片现场检测Pb2+
尽管基于脱氧核酶的Pb 2+检测取得了很大进展，但其在复杂介质中的适应性、灵敏度和可及性仍有待提高。为此，我们引入了克服这些障碍的新方法。首先，利用球形核酸（SNA）荧光探针（Au nanoparticle-DNAzyme probe）对Pb 2+进行特异性鉴定，并因其优异的抗核酸酶性而适用于复杂样品中Pb 2+的精确检测。二、Pb 2+的灵敏度通过使用具有目标识别精度的成簇的规则间隔短回文重复序列-Cas12a 来放大 SNA（信号探针）反式切割时的荧光信号，大大增强了检测能力，检测限低至 86 fM。第三，我们通过使用简单的检测设备（智能手机和便携式紫外灯）来增强具有仿生周期性排列的光子晶体芯片上的荧光，以实现 Pb 2+的现场检测，检测限低至 24 pM。基于上述努力，改进后的Pb 2+荧光传感器具有更高的灵敏度、更好的特异性、可及性、更少的样品消耗等优点。此外，它还可用于准确检测 Pb2+在复杂的生物或环境样品中，这对于广泛的应用具有很大的前景。
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