基于截短适配体-二硫化钼纳米片信号增强免标记荧光检测金刚烷胺
Detection of Amantadine by Label-free Fluorescence Method Based on Truncated Aptamer and Molybdenum Disulfide Nanosheet Signal Enhancement Strategy
兰艺凤 1侯博雅 1尉志文 1刘文 2张潮 1左雅慧 3贠克明1
作者信息
- 1. 山西医科大学法医学院,晋中 030600
- 2. 山西医科大学基础医学院,晋中 030600
- 3. 中国辐射防护研究院,太原 030000
- 折叠
摘要
金刚烷胺(AMD)可与其核酸适配体特异性结合,导致构象由单链转换成茎环结构.为避免多余的核苷酸影响适配体与靶标结合形成复杂的空间结构,本研究通过保留适当的茎环结构对AMD原适配体J进行截短优化,开发出新的截短适配体.与原适配体相比,截短适配体J-7与AMD的亲和力和特异性更好.以J-7为特异性识别元件,结合二硫化钼纳米片(MoS2Ns)信号放大技术构建了一种基于核酸适配体-MoS2Ns的超灵敏免标记荧光法用于AMD的检测,检出限为0.11 ng/mL.将本方法用于实际牛奶、酸奶及SD大鼠血清样品中AMD的检测,回收率为86.6%～108.2%.本研究为其它长序列适配体的截短提供了参考依据,也为食品中AMD残留的检测提供了更灵敏的检测方法.
Abstract
Amantadine(AMD)residue can accumulate in organisms through the food chain and cause serious harm to human body.AMD can specifically bind to AMD specific aptamer and cause its conformation to change from a random single strand to a stem-loop structure.To avoid the influence of excess nucleotides on binding of aptamer to AMD,the truncation of the AMD original aptamer J was optimized by retaining an appropriate stem-loop structure,and a new type of truncation aptamers was developed in this work.By comparing the truncated aptamer with the original aptamer,it was found that the truncated aptamer J-7 had better affinity and specificity with AMD.The detection limit of AMD was 0.11 ng/mL by using J-7 as specific recognition element and molybdenum disulfide nanosheet(MoS2Ns)as signal amplification element.The developed method base on truncated aptamer J-7 was used for detection of AMD in milk,yogurt and SD rat serum samples for the first time with recoveries of 86.6%-108.2%.This study provided a reference for truncating other long sequence aptamers and provided a more sensitive detection method for monitoring AMD residues in food.
关键词
截短适配体/二硫化钼纳米片/信号增强/金刚烷胺/免标记荧光法Key words
Truncated aptamer/Molybdenum disulfide nanosheet/Signal enhancement/Amantadine/Label-free fluorescence sensing system引用本文复制引用
基金项目
国家自然科学基金项目(82202083)
山西省自然科学基金项目(20210302124290)
出版年
2024
国家科技期刊平台
NSTL
万方数据