基于核酸适配体的荧光法检测水胺硫磷和丙溴磷

引用本文: 王丽, 叶华, 桑宏庆, 王丹丹. 基于核酸适配体的荧光法检测水胺硫磷和丙溴磷[J]. 分析化学, 2016, 44(5): 799-803. doi: 10.11895/j.issn.0253-3820.150601 shu

Citation: WANG Li, YE Hua, SANG Hong-Qing, WANG Dan-Dan. Aptamer-based Fluorescence Assay for Detection of Isocarbophos and Profenofos[J]. Chinese Journal of Analytical Chemistry, 2016, 44(5): 799-803. doi: 10.11895/j.issn.0253-3820.150601 shu

基于核酸适配体的荧光法检测水胺硫磷和丙溴磷

安徽科技学院食品药品学院, 凤阳 233100
基金项目:
本文系国家自然科学基金项目(No.31201369),安徽省高校优秀人才支持计划重点项目(No.gxyqZD2016219)和安徽科技学院自然科学研究项目(No.ZRC2014395)资助

摘要: 建立了基于适配体的农药水胺硫磷和丙溴磷的荧光检测方法。采用可特异性识别水胺硫磷和丙溴磷、且5'端标记荧光基团FAM的核酸适配体(F-ssDNA),与3'末端标记猝灭基团DABCYL的短链序列(Q-ssDNA)互补杂交形成双链结构,荧光基团的荧光被淬灭,荧光信号很弱;此时加入靶分子,特异性结合核酸适配体,引起互补短链序列从双链结构中解离,使适配体荧光信号增强,基于此可实现水胺硫磷、丙溴磷的定量检测。优化后的检测条件为:将终浓度为25 nmol/L F-ssDNA与50 nmol/L Q-ssDNA在25℃孵育20 min,使二者杂交形成双链适配体探针复合物,加入等体积的农药样品孵育60 min,然后检测体系的荧光信号变化值ΔI。在最佳条件下,ΔI与水胺硫磷和丙溴磷的浓度均在50-500μmol/L范围内呈线性关系。水胺硫磷的检出限(LOD,3σ)为11.4μmol/L,相对标准偏差(RSD)为5.8%(n=10);丙溴磷的检出限为14.0μmol/L,RSD为4.9%(n=10)。用于实际水样中两种农药的检测,加标回收率为85.8%~95.3%

关键词:

English

Aptamer-based Fluorescence Assay for Detection of Isocarbophos and Profenofos

College of Food and Drug, Anhui Science and Technology University, Fengyang 233100, China
Received Date: 28 July 2015
Revised Date: 13 January 2016
Fund Project:
This work was supported by the National Natural Science Foundation of China (No.31201369), the Key Project of College Excellent Talents Support Plan in Anhui Province (No. gxyqZD2016219) and the Natural Science Research Project of Anhui Science and Technology University (No. ZRC2014395).

Abstract: A fluorescence method for detection of isocarbophos and profenofos was established based on the specific recognition of aptamer. The aptamer recognizing isocarbophos and profenofos was labeled with fluorescent label FAM at 5' end (F-ssDNA). When it bound to complementary DNA chain labeled with quenching group DABCYL at 3' end (Q-ssDNA), the double-stranded structure would be formed and the fluorescence signals became very weak due to the effect of fluorescence resonance energy transfer. While the Q-ssDNA would be released from the double-stranded structure when the aptamer specifically recognized and bound the targets, and the fluorescence signals of the system would recover. Based on this, isocarbophos and profenofos could be quantitatively detected. The optimized experimental conditions were as follows:firstly, 25 nmol/L F-ssDNA and 50 nmol/L Q-ssDNA-2 were incubated for 20 min at 25℃, then an equal volume of pesticide was added and incubated for another 60 min, then the fluorescence intensity of the system was determined. Under the optimal conditions, the change of the fluorescence intensity of the system (ΔI) showed a linear relationship with the isocarbophos or profenofos concentration in the range of 50 to 500 μmol/L. The limit of detection (LOD, σ) was 11.4 μmol/L for isocarbophos with the relative standard deviation (RSD) of 5.8% (n=10). The LOD was 14.0 μmol/L for profenofos with RSD of 4.9% (n=10). The recovery ranged from 85.8% to 95.3% when this method was applied to detect isocarbophos and profenofos in real water samples.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

基于核酸适配体的荧光法检测水胺硫磷和丙溴磷

English

Aptamer-based Fluorescence Assay for Detection of Isocarbophos and Profenofos

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中国化学会秘书处

基于核酸适配体的荧光法检测水胺硫磷和丙溴磷

English

Aptamer-based Fluorescence Assay for Detection of Isocarbophos and Profenofos

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

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