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基于突变适体-智能手机比色法检测牛奶中的甲硝唑。

Based on mutated aptamer-smartphone colorimetric detection of metronidazole in milk.

作者信息

Zhang Sicheng, Qin Yadi, Yuan Jie, Wang Yu, Yao Jun, Zhang Minwei

机构信息

School of Pharmacy, Xinjiang Medical University, Urumqi, China.

School of Pharmacy, Xinjiang Second Medical College, Karamay, China.

出版信息

Front Bioeng Biotechnol. 2024 Aug 2;12:1444846. doi: 10.3389/fbioe.2024.1444846. eCollection 2024.

DOI:10.3389/fbioe.2024.1444846
PMID:39157440
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11327025/
Abstract

Excessive residue of metronidazole (MNZ) in food is harmful to the human body. There is an urgent demand to develop a portable tool for MNZ detection on-site. In this study, fifteen aptamers were prepared through targeted base mutation. Apt1-3 with the highest enrichment was chosen for further study. Its affinity was characterized by molecular docking simulation, AuNPs colorimetric assay, graphene oxide (GO) fluorescence assay, and exonuclease assay. Kd was determined by GO fluorescence assay (Kd: 92.60 ± 25.59 nM). Its specificity was also characterized by an exonuclease assay. A novel aptasensor was constructed by using the newly identified aptamer combined with the smartphone dark box. The principle of color change is caused by the aggregation state of AuNPs. Smartphones act as reading instruments. The detection can be completed in just a few seconds without the aid of instruments, achieving a detection limit of 0.15 nmol/mL and a range of 6.7-44.4 nmol/mL ( = 0.9810). Therefore, the constructed smartphone colorimetric sensor based on mutant aptamers has important applications in food detection.

摘要

食品中甲硝唑（MNZ）残留超标对人体有害。迫切需要开发一种用于现场检测MNZ的便携式工具。在本研究中，通过靶向碱基突变制备了15种适体。选择富集度最高的Apt1 - 3进行进一步研究。通过分子对接模拟、金纳米粒子比色法、氧化石墨烯（GO）荧光法和核酸外切酶法对其亲和力进行了表征。通过GO荧光法测定解离常数（Kd：92.60 ± 25.59 nM）。还通过核酸外切酶法对其特异性进行了表征。利用新鉴定的适体与智能手机暗盒相结合构建了一种新型适体传感器。颜色变化原理是由金纳米粒子的聚集状态引起的。智能手机作为读取仪器。无需借助仪器，仅需几秒钟即可完成检测，检测限为0.15 nmol/mL，线性范围为6.7 - 44.4 nmol/mL（R² = 0.9810）。因此，构建的基于突变适体的智能手机比色传感器在食品检测中具有重要应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/95590424f0e2/fbioe-12-1444846-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/1dc4614d706a/fbioe-12-1444846-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/0f917f4f491e/fbioe-12-1444846-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/2c8b79a3af6e/fbioe-12-1444846-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/581256dda725/fbioe-12-1444846-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/a62f82894471/fbioe-12-1444846-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/95590424f0e2/fbioe-12-1444846-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/1dc4614d706a/fbioe-12-1444846-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/0f917f4f491e/fbioe-12-1444846-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/2c8b79a3af6e/fbioe-12-1444846-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/581256dda725/fbioe-12-1444846-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/a62f82894471/fbioe-12-1444846-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2cd/11327025/95590424f0e2/fbioe-12-1444846-g006.jpg

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