基于核酸适配体功能化石墨纳米颗粒荧光探针的17β-雌二醇快速检测方法

doi:10.13278/j.cnki.jjuese.20180036

吉林大学学报(地球科学版) ›› 2019, Vol. 49 ›› Issue (4): 1137-1144.doi: 10.13278/j.cnki.jjuese.20180036

基于核酸适配体功能化石墨纳米颗粒荧光探针的17β-雌二醇快速检测方法

朴云仙¹, 祁小丽¹, 王湘¹, 康博泉¹, 史玉玺¹, 胡慧¹, 杨悦锁^1,2

1. 地下水资源与环境教育部重点实验室(吉林大学)/吉林大学新能源与环境学院, 长春 130021;
2. 区域污染环境生态修复教育部重点实验室(沈阳大学), 沈阳 110044

收稿日期:2018-03-02 出版日期:2019-07-26 发布日期:2019-07-26
作者简介:朴云仙(1979-),女,副教授,博士,主要从事纳米生物催化环境修复、环境监测和生物传感器方面的研究,E-mail:yxpiao@jlu.edu.cn
基金资助:
国家自然科学基金项目（51809111，41472237）

Rapid Detection of 17β-Estradiol Based on Fluorescent Probe of Functionalized Graphite Nanoparticle with Aptamers

Piao Yunxian¹, Qi Xiaoli¹, Wang Xiang¹, Kang Boquan¹, Shi Yuxi¹, Hu hui¹, Yang Yuesuo^1,2

1. Key Laboratory of Groundwater Resources and Environment(Jilin University), Ministry of Education/College of New Energy and Environment, Jilin University, Changchun 130021, China;
2. Key Laboratory of Eco-Restoration of Regional Contaminated Environment(Shenyang University), Ministry of Education, Shenyang 110044, China

Received:2018-03-02 Online:2019-07-26 Published:2019-07-26
Supported by:
Supported by National Natural Science Foundation of China (51809111, 41472237)

摘要/Abstract

摘要： 为构建一种能够简单、快速、特异性检测复杂环境水体中雌激素污染的方法，利用石墨纳米颗粒作为荧光淬灭剂、核酸适配体作为识别元素、1-芘丁酸N-羟基琥珀酰亚胺酯作为异型双功能交联剂，构建了一种新型纳米荧光探针；并探究了构建荧光探针时核酸适配体初始投加量和荧光探针投加量对雌二醇检测的影响及最佳实验条件下检测雌二醇的效果和特异性。实验结果表明：核酸适配体能成功修饰在石墨纳米颗粒表面形成的稳定荧光探针；构建荧光探针时核酸适配体的最佳初始投加量为1.0 μmol/L；检测雌二醇时，荧光探针的最佳投加量为4 μg/mL；最佳实验条件下，相对荧光强度与雌二醇的质量浓度在50~800 ng/mL范围内成正比，最低检测限为34.5 ng/mL，且该荧光探针能实现对雌二醇的简单、快速、特异性检测。

关键词: 石墨纳米颗粒, 核酸适配体, 17β-雌二醇, 探针, 快速检测

Abstract: In order to establish a simple, rapid and specific method for detecting 17β-estradiol in complex environmental water, we prepared a novel fluorescence probe by using graphite nanoparticles(GN) as excellent quencher, aptamers as recognition elements,and 1-pyrenebutyric acid N-hydroxysuccinimide ester as heterobifunctional crosslinking reagent. We studied the effects of initial concentration of aptamer on the construction of aptamer probe, the change of aptamer probe's concentration on fluorescence signals, and the optimal conditions for 17β-estradiol detection. The results showed that the aptamers were successfully anchored on the surface of GN, and the fluorescence aptamer probe was well synthesized. The optimum initial concentration of aptamer for the preparation of fluorescence probe was 1.0 μmol/L; the optimum concentration of the fluorescent probe for analysis was 4 μg/mL. Under the optimal condition, the relative fluorescence intensity was proportional to the concentration of 17β-estradiol ranging from 50 to 800 ng/mL, and the limit of detection was 34.5 ng/mL. This method is simple, quick and specific for the detection of 17β-estradiol.

Key words: graphite nanoparticle, aptamer, 17β-estradiol, probe, rapid detection

中图分类号:

X502

朴云仙, 祁小丽, 王湘, 康博泉, 史玉玺, 胡慧, 杨悦锁. 基于核酸适配体功能化石墨纳米颗粒荧光探针的17β-雌二醇快速检测方法[J]. 吉林大学学报(地球科学版), 2019, 49(4): 1137-1144.

Piao Yunxian, Qi Xiaoli, Wang Xiang, Kang Boquan, Shi Yuxi, Hu hui, Yang Yuesuo. Rapid Detection of 17β-Estradiol Based on Fluorescent Probe of Functionalized Graphite Nanoparticle with Aptamers[J]. Journal of Jilin University(Earth Science Edition), 2019, 49(4): 1137-1144.

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基于核酸适配体功能化石墨纳米颗粒荧光探针的17β-雌二醇快速检测方法

Rapid Detection of 17β-Estradiol Based on Fluorescent Probe of Functionalized Graphite Nanoparticle with Aptamers

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