title: A triple-amplification SPR electrochemiluminescence assay for chloramphenicol based on polymer enzyme-linked nanotracers and exonuclease-assisted target recycling.
abstract: The present study aimed to explore a novel triple-amplification electrochemiluminescence (ECL) assay for detecting of chloramphenicol (CAP). This strategy was based on single-stranded DNA-binding protein (SSB) and horseradish peroxidase (HRP) enzyme-linked polymer (EnVision reagent, EV) labeled on Au nanoparticles (EV-Au-SSB) as nanotracer and exonuclease-assisted target recycling. The composite probes were prepared via immunoreactions between the CdS nanocrystal (CdS NC)-functionalized partial complementary DNA and aptamer (CdSNCs/Apt-ssDNA1) as capture probes, and EV-Au-SSB as nanotracer. When the composite probe solution co-existed with CAP and Exo I, the aptamer on the capture probes preferentially combined with CAP, and then CAP-Apt and nanotracer complex were released into the solution. Subsequently, Exo I in the solution could further digest the CAP-Apt from the 3'-end of the aptamer and release CAP, which could participate in further reaction with the probes. It was worth mentioning that EV contained a large number of HRPs on its dendritic chain. In the EV-Au-SSB, Au could enhance ECL intensity of CdS NCs by surface plasmon resonance. What's more, HRPs on EV could catalyze the reaction of H2O2, which could obviously enhance ECL intensity of CdS NCs. This study demonstrated excellent performance of the triple-amplification ECL assay, which makes this aptasensor system suitable and promising for the practical application of CAP residues in fish samples. Moreover, the assay might provide a promising avenue to develop efficient aptasensors to determine small-molecule harmful substances in environmental monitoring and food safety.
authors.affiliation: State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, PR China.
authors.forename: Yang-Bao
authors.initials: YB
authors.lastname: Miao
authors.affiliation: Key Laboratory of Asymmetric Synthesis and Chirotechnology of Sichuan Province, Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu 610041, PR China; University of Chinese Academy of Sciences, Beijing 10049, PR China.
authors.forename: Hong-Xia
authors.initials: HX
authors.lastname: Ren
authors.affiliation: State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, PR China. Electronic address: ganning@nbu.edu.cn.
authors.forename: Ning
authors.initials: N
authors.lastname: Gan
authors.affiliation: State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, PR China. Electronic address: zhouyou@nbu.edu.cn.
authors.forename: You
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, PR China.
authors.forename: Yuting
authors.initials: Y
authors.lastname: Cao
authors.affiliation: State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, PR China. Electronic address: litianhua@nbu.edu.cn.
authors.forename: Tianhua
authors.initials: T
authors.lastname: Li
authors.affiliation: Faculty of Food Science and Engineering, Nanjing University of Finance and Economics, Nanjing 210000, PR China.
authors.forename: Yinji
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000900:Anti-Bacterial Agents / Q000032:analysis*; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D019187:Cadmium Compounds / Q000737:chemistry; D002701:Chloramphenicol / Q000032:analysis*; D004268:DNA-Binding Proteins / Q000737:chemistry; D055664:Electrochemical Techniques / Q000379:methods*; D004785:Environmental Pollutants / Q000032:analysis; D006046:Gold / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry; D006861:Hydrogen Peroxide / Q000737:chemistry; D057230:Limit of Detection; D008163:Luminescent Measurements / Q000379:methods; D053758:Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D013440:Sulfides / Q000737:chemistry
chemical_list: D000900:Anti-Bacterial Agents; D052157:Aptamers, Nucleotide; D019187:Cadmium Compounds; D004268:DNA-Binding Proteins; D004785:Environmental Pollutants; D013440:Sulfides; C034939:cadmium sulfide; D002701:Chloramphenicol; D006046:Gold; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase
keywords: Chloramphenicol; Horseradish peroxidase enzyme-linked polymer; Nanotracer; Triple-amplification electrochemiluminescence assay; single-stranded DNA-binding protein