DNAzyme-Au nanoprobe coupled with graphene-oxide–loaded hybridization chain reaction signal amplification for fluorometric determination of alkaline phosphatase,Microchimica Acta

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DNAzyme-Au nanoprobe coupled with graphene-oxide–loaded hybridization chain reaction signal amplification for fluorometric determination of alkaline phosphatase
Microchimica Acta ( IF 5.3 ) Pub Date : 2021-01-01 , DOI: 10.1007/s00604-020-04681-1
Zhengxian Lv ₁ , Qiuquan Wang ₁ , Minghui Yang ₂

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A sensing platform is presented for the determination of alkaline phosphatase (ALP) activity based on the cooperation of DNAzyme-Au spherical nucleic acid nanoprobe with the graphene-oxide–loaded hybridization chain reaction (HCR/GO) system to achieve good detection sensitivity and specificity. This assay takes advantage of the strong affinity of pyrophosphate (PPi) to Cu 2+ ions and the fact that ALP can hydrolyze pyrophosphate (PPi) to release free Cu 2+ ions. In the presence of ALP, the released Cu 2+ can promote the Cu 2+ -dependent DNAzyme to cleave the substrate that generates a shorter DNA fragment, which is responsible for further triggering the HCR/GO system to form a long fluorescence dsDNA and thereby giving an amplified fluorescence signal. Linear calibration range was obtained from 0.2 to 10 U L −1 , and the limit of detection (LOD) is about 0.14 U L −1 . The feasibility of the proposed method was validated by spiking ALP standards in bovine serum. The recovery ranged from 97.2 to 104.6%, and a coefficient of variation (CV) of less than 8% ( n = 3) was obtained. This assay strategy was also applied to evaluate the ALP inhibitor efficiency, which indicates that the assay has potential for drug screening. Graphical abstract

中文翻译：

DNAzyme-Au 纳米探针结合氧化石墨烯杂交链反应信号放大用于碱性磷酸酶的荧光测定

基于 DNAzyme-Au 球形核酸纳米探针与氧化石墨烯杂交链反应 (HCR/GO) 系统的合作，提出了一种用于测定碱性磷酸酶 (ALP) 活性的传感平台，以实现良好的检测灵敏度和特异性. 该测定利用焦磷酸盐 (PPi) 对 Cu 2+ 离子的强亲和力以及 ALP 可以水解焦磷酸盐 (PPi) 以释放游离 Cu 2+ 离子的事实。在 ALP 存在下，释放的 Cu 2+ 可以促进 Cu 2+ 依赖性 DNAzyme 切割底物，产生较短的 DNA 片段，这负责进一步触发 HCR/GO 系统形成长荧光 dsDNA，从而产生放大的荧光信号。线性校准范围从 0.2 到 10 U L -1 ，并且检测限(LOD)约为0.14 U L -1 。通过在牛血清中添加 ALP 标准品来验证所提出方法的可行性。回收率范围为 97.2% 至 104.6%，变异系数 (CV) 小于 8% (n = 3)。该测定策略还用于评估 ALP 抑制剂的效率，这表明该测定具有药物筛选的潜力。图形概要

更新日期：2021-01-01

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