A magnetic bead-based dual-aptamer sandwich assay for quantitative detection of ciprofloxacin using CRISPR/Cas12a

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 palin...

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Main Authors: Fangyue Guo, Jianghao Li, Peizhi Ma, Mengying Liu, Jing Wu, Hai Qu, Yehuan Zheng, Mengying Wang, Seyed Sepehr Marashi, Zhijian Zhang, Shanfeng Zhang, Guangyu Fu, Pei Li
Format: Article
Language:English
Published: Elsevier 2025-02-01
Series:Molecular and Cellular Probes
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Online Access:http://www.sciencedirect.com/science/article/pii/S0890850824000501
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author Fangyue Guo
Jianghao Li
Peizhi Ma
Mengying Liu
Jing Wu
Hai Qu
Yehuan Zheng
Mengying Wang
Seyed Sepehr Marashi
Zhijian Zhang
Shanfeng Zhang
Guangyu Fu
Pei Li
author_facet Fangyue Guo
Jianghao Li
Peizhi Ma
Mengying Liu
Jing Wu
Hai Qu
Yehuan Zheng
Mengying Wang
Seyed Sepehr Marashi
Zhijian Zhang
Shanfeng Zhang
Guangyu Fu
Pei Li
author_sort Fangyue Guo
collection DOAJ
description 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.
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spelling doaj-art-fb5eb7825bd64cf79b9ae53f03c4db612025-08-20T02:13:52ZengElsevierMolecular and Cellular Probes0890-85082025-02-017910199810.1016/j.mcp.2024.101998A magnetic bead-based dual-aptamer sandwich assay for quantitative detection of ciprofloxacin using CRISPR/Cas12aFangyue Guo0Jianghao Li1Peizhi Ma2Mengying Liu3Jing Wu4Hai Qu5Yehuan Zheng6Mengying Wang7Seyed Sepehr Marashi8Zhijian Zhang9Shanfeng Zhang10Guangyu Fu11Pei Li12School 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 ChinaR & D Center, Autobio Diagnostics Co., Ltd, Zhengzhou, Henan, 450016, PR ChinaSchool 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 ChinaSchool 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 ChinaSchool 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 ChinaR & D Center, Autobio Diagnostics Co., Ltd, Zhengzhou, Henan, 450016, PR ChinaSchool of Basic Medical Sciences, Xi'an Jiaotong University, Xian, 710061, PR ChinaState Key Laboratory of Biotherapy, Sichuan University, Chengdu, 610041, PR ChinaCollege of International Education, Zhengzhou University, Zhengzhou, Henan, 450001, PR ChinaSchool of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan, 450001, PR ChinaSchool 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; Corresponding author. School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan, 450001, PR China.R & D Center, Autobio Diagnostics Co., Ltd, Zhengzhou, Henan, 450016, PR China; Corresponding author.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; Corresponding author. School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan, 450001, PR China.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.http://www.sciencedirect.com/science/article/pii/S0890850824000501Dual-aptamer sandwich assayCiprofloxacinCRISPR/Cas12aDetection
spellingShingle Fangyue Guo
Jianghao Li
Peizhi Ma
Mengying Liu
Jing Wu
Hai Qu
Yehuan Zheng
Mengying Wang
Seyed Sepehr Marashi
Zhijian Zhang
Shanfeng Zhang
Guangyu Fu
Pei Li
A magnetic bead-based dual-aptamer sandwich assay for quantitative detection of ciprofloxacin using CRISPR/Cas12a
Molecular and Cellular Probes
Dual-aptamer sandwich assay
Ciprofloxacin
CRISPR/Cas12a
Detection
title A magnetic bead-based dual-aptamer sandwich assay for quantitative detection of ciprofloxacin using CRISPR/Cas12a
title_full A magnetic bead-based dual-aptamer sandwich assay for quantitative detection of ciprofloxacin using CRISPR/Cas12a
title_fullStr A magnetic bead-based dual-aptamer sandwich assay for quantitative detection of ciprofloxacin using CRISPR/Cas12a
title_full_unstemmed A magnetic bead-based dual-aptamer sandwich assay for quantitative detection of ciprofloxacin using CRISPR/Cas12a
title_short A magnetic bead-based dual-aptamer sandwich assay for quantitative detection of ciprofloxacin using CRISPR/Cas12a
title_sort magnetic bead based dual aptamer sandwich assay for quantitative detection of ciprofloxacin using crispr cas12a
topic Dual-aptamer sandwich assay
Ciprofloxacin
CRISPR/Cas12a
Detection
url http://www.sciencedirect.com/science/article/pii/S0890850824000501
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