荧光核酸适配体功能化氧化石墨烯生物传感器用于快速检测氯霉素
Fluorescent Aptamer-functionalized Graphene Oxide Biosensor for Rapid Detection of Chloramphenicol
Received date: 2018-10-17
Online published: 2019-01-17
Supported by
Project supported by the Scientific Research Fund of Hunan Provincial Education Department (No. 17C0033).
开发了一种无标记和快速的检测方法基于氧化石墨烯（GO）和荧光功能性G-四聚体探针（FGP），可用于定量检测氯霉素（CAP）.FGP由氯霉素核酸适配体和富含G碱基的核酸序列组成.核酸适配体用于结合CAP，并且由富含G碱基的核酸序列在K+，Na+离子的作用下形成的G-四聚体，然后与硫磺素T（ThT）结合后用作信号分子.在没有CAP的情况下，FGP通过π-π堆积相互作用被吸附到GO的表面上，阻碍了G-四聚体的形成导致溶液中的荧光强度低.在加入CAP时，FGP的核酸适配体部分可识别并结合CAP以形成复合物，导致其从GO解吸.因此，游离的富含G的碱基序列可以形成G-四聚体结构并与ThT结合，导致溶液的荧光强度增加.我们观察到荧光强度增加与CAP浓度在2~20 nmol/L范围内呈线性关系，检测限为1.45 nmol/L.此外，该检测系统用于检测加标牛奶中的CAP，回收率在93.2%~103.3%之间.这些结果表明，开发的方法可用于有效检测实际样品中的CAP.
卢静荷 , 谭淑珍 , 朱雨清 , 李伟 , 陈天啸 , 王瑶 , 刘陈 . 荧光核酸适配体功能化氧化石墨烯生物传感器用于快速检测氯霉素[J]. 化学学报, 2019 , 77(3) : 253 -256 . DOI: 10.6023/A18100433
A label free and rapid fluorescent method for quantitative detection of chloramphenicol (CAP) based on graphene oxide (GO) fluorescence functional G-quadruplex probe (FGP) was developed. The FGP consisted of a choramphenicol aptamer and a G-rich sequence. The aptamer was used to bind CAP and the G-quadruplex formed by G-rich sequence was employed as a signal reporter after binding to Thioflavin T (ThT). In the absence of CAP, the FGP was absorbed onto the surface of GO through π-π stacking interactions, which restrained the G-rich sequence to form a G-quadruplex structure. Thus, the fluorescent intensity of background was low. In the addition of the CAP, the aptamer part of FGP could recognize and bind CAP to form a target-FGP complex, which led to the desorption of the complex from GO. Therefore, the free G-rich sequence could form G-quadruplex structure and bind to ThT, resulting a increase in the fluorescence intensity of the solution. We observed that the fluorescence increasement of the sensing platform had a linear relationship with the concentrations of CAP in the range of 2~20 nmol/L, and the limit of detection was 1.45 nmol/L. Besides, this detection system was applied for detecting CAP in the spiked milk, the recovery rate was between 93.2%~103.3%. These results indicated that this developed method can be used to efficiently recognize CAP in real samples.
Key words： aptamer; fluorescence; chloramphenicol; G-quadruplex; graphene oxide
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