Guo Fangyue, Li Jianghao, Ma Peizhi, Liu Mengying, Wu Jing, Qu Hai, Zheng Yehuan, Wang Mengying, Marashi Seyed Sepehr, Zhang Zhijian, Zhang Shanfeng, Fu Guangyu, Li Pei
School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan, 450001, PR China; School of Basic Medical Sciences Innovation and Entrepreneurship Base for College Students, Zhengzhou University, Zhengzhou, Henan, 450001, PR China; Department of the First Clinical Medicine, Zhengzhou University, Zhengzhou, Henan, 450052, PR China.
R & D Center, Autobio Diagnostics Co., Ltd, Zhengzhou, Henan, 450016, PR China.
Mol Cell Probes. 2025 Feb;79:101998. doi: 10.1016/j.mcp.2024.101998. Epub 2024 Dec 24.
Ciprofloxacin (CIP) is a broad-spectrum fluoroquinolone antibiotic, and its excessive residues in food and water sources pose potential risks to human health. Therefore, there is a need for a rapid and convenient method for its accurate quantification. The clustered regularly interspaced short palindromic repeat (CRISPR)/Cas12a system has gained extensive application in signal detection and amplification due to the trans-cleavage activity of Cas12a. In this study, we devised a novel magnetic bead-based dual sandwich aptamer coupled with a CRISPR/Cas12a system for the precise quantification of CIP in milk, river water, and honey. Through the incorporation of a magnetic bead-based dual aptamer sandwich approach, the concentration of CIP in the samples was pre-enriched. Additionally, by optimizing the Fluorescence-Quencher (F-Q) probe concentration, detection aptamer (APTd) concentration, and assay duration, the limit of blank (LOB) of the system was determined as 362 nM, while the limit of detection (LOD) was determined as 403 nM. This enabled the accurate quantification of CIP within the linear range of 0.5 μM to 0.2 mM with high specificity. Moreover, the performance of this detection method was comparable to that of high-performance liquid chromatography (HPLC) in river water, milk, and honey samples.
环丙沙星（CIP）是一种广谱氟喹诺酮类抗生素，其在食物和水源中的过量残留对人类健康构成潜在风险。因此，需要一种快速便捷的方法对其进行准确定量。由于Cas12a的反式切割活性，成簇规律间隔短回文重复序列（CRISPR）/Cas12a系统在信号检测和放大方面得到了广泛应用。在本研究中，我们设计了一种基于磁珠的新型双夹心适配体与CRISPR/Cas12a系统相结合的方法，用于精确测定牛奶、河水和蜂蜜中的CIP。通过采用基于磁珠的双适配体夹心方法，对样品中CIP的浓度进行了预富集。此外，通过优化荧光淬灭（F-Q）探针浓度、检测适配体（APTd）浓度和检测时间，确定该系统的空白限（LOB）为362 nM，检测限（LOD）为403 nM。这使得能够在0.5 μM至0.2 mM的线性范围内以高特异性对CIP进行准确定量。此外，该检测方法在河水、牛奶和蜂蜜样品中的性能与高效液相色谱（HPLC）相当。
