In vitro selection and engineering azithromycin-specific aptamers and construction of a ratiometric fluorescent aptasensor for sensitive detection of azithromycin

阿奇霉素 适体 指数富集配体系统进化 化学 离解常数 色谱法 检出限 组合化学 生物 分子生物学 生物化学 核糖核酸 抗生素 基因 受体
作者
Tianyu Huang,Xin Chen,Jinri Chen,Yuting Zhang,Xiaoli Wang,Zhimeng Wu,Nandi Zhou
出处
期刊:Sensors and Actuators B-chemical [Elsevier BV]
卷期号:411: 135789-135789 被引量:6
标识
DOI:10.1016/j.snb.2024.135789
摘要

Azithromycin can effectively inhibit bacterial protein synthesis and reduce biofilm formation and quorum sensing, and thus has been preferentially used in the treatment of various infections. However, azithromycin is also among the most highly concentrated antibiotics present in wastewater due to the inappropriate use and treatment. In order to sensitively and rapidly monitor the azithromycin residues, aptamers specific to azithromycin were selected through 12 rounds of screening by using magnetic beads-SELEX (MB-SELEX). The screened aptamers were assessed for their affinity and specificity using fluorescent assay. Apt3 emerged as the optimal candidate aptamer with the dissociation constant (Kd) of 235.07 nM. After secondary structure analysis and molecular docking, Apt3 was systematically truncated and subjected to single-base mutations. The optimal aptamer Apt3–27 T was identified, exhibiting the Kd value of 215.84 nM and the improved specificity compared to the original aptamer. Then Apt3–27 T was utilized to construct a ratiometric fluorescent aptasensor, which exhibited the linear dynamic range of 25–400 nM and low detection limit of 9.78 nM. More importantly, the ratiometric fluorescence mechanism further improved the specificity of the biosensor, which finally can discriminate azithromycin from structurally similar erythromycin. Evaluation experiments using actual samples spiked with azithromycin showed the recovery ranging from 97.1% to 106% and the relative standard deviation ranging from 4.29% to 8.61%. The screened aptamer and constructed aptasensor offer the advantages of high affinity, high effectiveness and ease of use in actual sample detection, thereby displaying promising applications in the monitoring of azithromycin residues.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
KK完成签到,获得积分10
1秒前
蓝桉完成签到,获得积分10
2秒前
zhouxuefeng发布了新的文献求助10
2秒前
3秒前
kanwenxian完成签到,获得积分20
3秒前
期刊发布了新的文献求助50
3秒前
共享精神应助小夫同学采纳,获得10
4秒前
4秒前
小蘑菇应助树袋采纳,获得10
4秒前
科研通AI5应助xn201120采纳,获得10
5秒前
5秒前
hh完成签到,获得积分20
6秒前
王彬完成签到,获得积分10
6秒前
晚来天欲雪完成签到,获得积分20
8秒前
Lc应助蓝桉采纳,获得20
8秒前
13秒前
XXXXL完成签到,获得积分10
15秒前
麦苗果果发布了新的文献求助10
17秒前
小夫同学发布了新的文献求助10
17秒前
18秒前
英姑应助谦让小松鼠采纳,获得10
18秒前
BKEL完成签到,获得积分10
21秒前
21秒前
lalala驳回了SciGPT应助
23秒前
kanwenxian发布了新的文献求助10
24秒前
今后应助解语花采纳,获得10
25秒前
七慕凉应助解语花采纳,获得10
25秒前
FashionBoy应助pineapple yang采纳,获得20
25秒前
麦苗果果完成签到,获得积分10
25秒前
Irene完成签到,获得积分10
26秒前
小二郎应助蓁66采纳,获得10
27秒前
27秒前
Hello应助陈曦采纳,获得10
27秒前
领导范儿应助hh采纳,获得10
28秒前
29秒前
艺涵发布了新的文献求助10
31秒前
孙燕应助闪闪泥猴桃采纳,获得30
32秒前
34秒前
34秒前
35秒前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
‘Unruly’ Children: Historical Fieldnotes and Learning Morality in a Taiwan Village (New Departures in Anthropology) 400
Indomethacinのヒトにおける経皮吸収 400
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
基于可调谐半导体激光吸收光谱技术泄漏气体检测系统的研究 350
Robot-supported joining of reinforcement textiles with one-sided sewing heads 320
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3989550
求助须知:如何正确求助?哪些是违规求助? 3531774
关于积分的说明 11254747
捐赠科研通 3270278
什么是DOI,文献DOI怎么找? 1804966
邀请新用户注册赠送积分活动 882125
科研通“疑难数据库(出版商)”最低求助积分说明 809176