Fluorescence immunoassay based on the enzyme cleaving ss-DNA to regulate the synthesis of histone-ds-poly(AT) templated copper nanoparticles
Fluorescence immunoassay based on the enzyme cleaving ss-DNA to regulate the synthesis of histone-ds-poly(AT) templated copper nanoparticles
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基于酶切 ss-DNA 的荧光免疫分析调控组蛋白-ds-poly(AT) 模板铜纳米粒子的合成
DOI：
10.1039/c8nr06175k
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发表时间：
2018
期刊：
影响因子：
6.7
通讯作者：
Xiong Yonghua
中科院分区：
文献类型：
--
作者：
Xiong Ying;Gao Bao;Wu Kesheng;Wu Yunqing;Chai Yinjiao;Huang Xiaolin;Xiong Yonghua
在这里，我们首次报道了一种新的竞争荧光免疫分析法，用于超灵敏检测黄曲霉毒素B1 (AFB1)，使用组蛋白聚（AT）模板铜纳米粒子（His-pAT CuNPs）作为荧光指示剂。在这个免疫分析中，葡萄糖氧化酶（Gox）被用作竞争抗原的载体，催化葡萄糖生成过氧化氢（H2O2）。利用Fenton试剂将H2O2转化为羟基自由基，进一步调控His-pAT CuNPs的荧光信号。由于ss-DNA对羟基自由基具有极高的敏感性，该荧光免疫法对AFB1的动态线性检测范围为0.46 pg mL - 1 ~ 400 pg mL - 1，具有良好的一半最大抑制浓度，检测限分别为6.13和0.15 pg mL - 1。结果表明，AFB1加标玉米样品的平均加样回收率分别为96.87% ~ 100.73%和96.67% ~ 114.92%。采用超高效液相色谱-荧光检测法进一步验证了该方法的可靠性。总之，这项工作为真菌毒素或其他污染物的定量检测提供了一种具有高灵敏度和鲁棒性的新型筛选策略，用于食品安全和临床诊断。
Herein, for the first time we report a novel competitive fluorescence immunoassay for the ultrasensitive detection of aflatoxin B1 (AFB1) using histone-ds-poly(AT) templated copper nanoparticles (His-pAT CuNPs) as the fluorescent indicator. In this immunoassay, glucose oxidase (Gox) was used as the carrier of the competing antigen to catalyze the formation of hydrogen peroxide (H2O2) from glucose. H2O2 was converted to a hydroxyl radical using Fenton's reagent, which further regulated the fluorescence signals of His-pAT CuNPs. Owing to the ultrahigh sensitivity of the ss-DNA to the hydroxyl radical, the proposed fluorescence immunoassay exhibited a favorable dynamic linear detection of AFB1 ranging from 0.46 pg mL−1 to 400 pg mL−1 with an good half maximal inhibitory concentration and limit of detection of 6.13 and 0.15 pg mL−1, respectively. The intra- and inter-assay showed that the average recoveries for AFB1 spiked corn samples ranged from 96.87% to 100.73% and 96.67% to 114.92%, respectively. The reliability of this method was further confirmed by adopting ultra-performance liquid chromatography coupled with the fluorescence detector method. In summary, this work offers a novel screening strategy with high sensitivity and robustness for the quantitative detection of mycotoxins or other pollutants for food safety and clinical diagnosis.