Structure-switching fluorescence aptasensor for sensitive detection of chloramphenicol,Microchimica Acta

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Structure-switching fluorescence aptasensor for sensitive detection of chloramphenicol
Microchimica Acta ( IF 5.3 ) Pub Date : 2020-08-19 , DOI: 10.1007/s00604-020-04471-9
Pengfei Ma _{1,

2,

3} , Yuhan Sun _{1,

2,

3} , Imran Mahmood Khan _{1,

2,

3} , QianHui Gu _{1,

2,

3,

4} , Lin Yue _{1,

2,

3} , Zhouping Wang _{1,

2,

3,

5,

6}

Affiliation

The performance of chloramphenicol aptamer, including binding thermodynamics, structure switching, and binding domain, was investigated by isothermal titration calorimetry, circular dichroism, and molecular docking. Then, a new fluorescence aptasensor was developed with signal amplification mediated by exonuclease I-catalyzed reaction and hybridization chain reaction (HCR) for chloramphenicol detection. In this system, the aptamer-binding domain is blocked by the initiator of HCR, the aptamer undergoes structure switching in the presence of chloramphenicol, and DNA dissociation occurs. The released aptamer is subsequently recognized and cleaved by Exo I to set free chloramphenicol. With the Exo I-assisted chloramphenicol recycling, an increasing number of initiators were exposed from the digestion of the initiator-aptamer complex. Then, the chain-like assembly of FAM labeled H1 and H2 through HCR was triggered by the initiator, generating a long DNA polymer. Under optimum conditions, the aptasensor exhibited a log-linear range from 0.001 to 100 nM of chloramphenicol and a detection limit of 0.3 pM. Additionally, the designed biosensing platform was applied to determine chloramphenicol in milk and lake water with high accuracy. The current approach provides a new avenue to develop sensitive aptasensors with the assistance of binding mechanism between aptamer and target compounds. Graphical abstract Graphical abstract

中文翻译：

用于灵敏检测氯霉素的结构转换荧光适体传感器

通过等温滴定量热法、圆二色性和分子对接研究了氯霉素适体的性能，包括结合热力学、结构转换和结合域。然后，开发了一种新的荧光适体传感器，其信号放大由外切核酸酶 I 催化反应和杂交链反应 (HCR) 介导，用于检测氯霉素。在该系统中，适体结合域被 HCR 的引发剂阻断，适体在氯霉素存在下进行结构转换，DNA 发生解离。释放的适体随后被 Exo I 识别并裂解以释放氯霉素。随着 Exo I 辅助氯霉素回收，越来越多的引发剂从引发剂-适体复合物的消化中暴露出来。然后，FAM 通过 HCR 标记 H1 和 H2 的链状组装由引发剂触发，生成长 DNA 聚合物。在最佳条件下，适体传感器表现出 0.001 至 100 nM 氯霉素的对数线性范围和 0.3 pM 的检测限。此外，设计的生物传感平台还用于高精度测定牛奶和湖水中的氯霉素。目前的方法为利用适体和目标化合物之间的结合机制开发灵敏的适体传感器提供了新的途径。图形摘要图形摘要设计的生物传感平台用于高精度测定牛奶和湖水中的氯霉素。目前的方法为利用适体和目标化合物之间的结合机制开发灵敏的适体传感器提供了新的途径。图形摘要图形摘要设计的生物传感平台用于高精度测定牛奶和湖水中的氯霉素。目前的方法为利用适体和目标化合物之间的结合机制开发灵敏的适体传感器提供了新的途径。图形摘要图形摘要

更新日期：2020-08-19

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