{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T05:01:02Z","timestamp":1781586062907,"version":"3.54.5"},"reference-count":37,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2022,10,13]],"date-time":"2022-10-13T00:00:00Z","timestamp":1665619200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Natural Science Foundation of Hebei Province","award":["C2020204130"],"award-info":[{"award-number":["C2020204130"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Foods"],"abstract":"<jats:p>The residues of aminoglycosides in foods of animal origin are a potential risk to consumers. There have been some immunoassays reported for the screening of aminoglycoside residues, but the method showing the broadest detection spectrum can only be used to detect two drugs. This is because a broad specific recognition reagent is not available. In the present study, the receptor of aminoglycosides (ribosomal protein S12 of Lysinibacillussphaericus) was expressed, and its affinities and recognition mechanisms for 10 aminoglycosides were studied by using surface plasmon resonance and molecular docking, respectively. Then the receptor was used as a recognition reagent to develop a fluorescence polarization assay on a 96-well microplate for the detection of the 10 drugs in pork muscle samples. The limits of detection for the 10 drugs ranged from 5.25 to 30.25 ng\/g. The sensitivities for the 10 drugs were generally consistent with their respective receptor affinities and binding energies. After comprehensive comparison, the method performances were better than all the previously reported immunoassays for aminoglycosides. This is the first study reporting the recognition mechanisms of ribosomal protein S12 of Lysinibacillussphaericus for 10 aminoglycosides and the use of it as a recognition reagent to develop a pseudo-immunoassay for the multi-determination of aminoglycosides in food samples.<\/jats:p>","DOI":"10.3390\/foods11203196","type":"journal-article","created":{"date-parts":[[2022,10,13]],"date-time":"2022-10-13T20:55:11Z","timestamp":1665694511000},"page":"3196","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Development of a Fluorescence Polarization Assay for Multi-Determination of 10 Aminoglycosides in Pork Muscle Sample Based on Ribosomal Protein S12 and Studying Its Recognition Mechanism"],"prefix":"10.3390","volume":"11","author":[{"given":"Wanqiu","family":"Xia","sequence":"first","affiliation":[{"name":"College of Veterinary Medicine, Hebei Agricultural University, Baoding 071000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lei","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Hebei Agricultural University, Baoding 071000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianping","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Hebei Agricultural University, Baoding 071000, China"},{"name":"Veterinary Biological Technology Innovation Center of Hebei Province, Baoding 071000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1421","DOI":"10.1136\/bmj.305.6866.1421","article-title":"ABC of monitoring drug therapy, aminoglycoside antibiotics","volume":"305","author":"Aronson","year":"1992","journal-title":"BMJ"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"119","DOI":"10.2174\/138161207779313731","article-title":"Aminoglycoside-induced ototoxicity","volume":"3","author":"Selimoglu","year":"2007","journal-title":"Curr. Pharm. Des."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.aca.2015.06.038","article-title":"Challenges in the determination of aminoglycoside antibiotics, a review","volume":"890","author":"Farouk","year":"2015","journal-title":"Anal. Chim. Acta"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"145","DOI":"10.7324\/JAPS.2019.91118","article-title":"Challenges in the development of analytical test procedure for aminoglycosides: A critical review","volume":"9","author":"Hari","year":"2019","journal-title":"J. Appl. Pharm. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2944","DOI":"10.1021\/jf703602b","article-title":"Rapid enzyme-linked immunosorbent assay and colloidal gold immunoassay for kanamycin and tobramycin in swine tissues","volume":"56","author":"Chen","year":"2008","journal-title":"J. Agric. Food Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"964","DOI":"10.1039\/b418107g","article-title":"Enzyme immunoassays for the analysis of streptomycin in milk, serum and water: Development and assessment of a polyclonal antiserum and assay procedures using novel streptomycin derivatives","volume":"130","author":"Abuknesha","year":"2005","journal-title":"Analyst"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.jpba.2012.11.014","article-title":"Development of generic immunoassay for the detection of a series of aminoglycosides with 6\u2032-OH group for the treatment of genetic diseases in biological samples","volume":"75","author":"Shalev","year":"2013","journal-title":"J. Pharmaceut. Biomed."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1016\/j.bios.2013.04.027","article-title":"Development of a novel bead-based 96-well filtration plate competitive immunoassay for the detection of Gentamycin","volume":"49","author":"Ho","year":"2013","journal-title":"Biosens. Bioelectron."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"3108","DOI":"10.1021\/jf405379r","article-title":"Production of monoclonal antibody and development of a new immunoassay for apramycin in food","volume":"62","author":"Xu","year":"2014","journal-title":"J. Agric. Food Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2066","DOI":"10.1007\/s12161-018-1185-2","article-title":"An enhanced direct competitive immunoassay for the detection of kanamycin and tobramycin in milk using multienzyme-particle amplification","volume":"11","author":"Jiang","year":"2018","journal-title":"Food Anal. Methods"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1039\/C8AY02006J","article-title":"Visual dual dot immunoassay for the simultaneous detection of kanamycin and streptomycin in milk","volume":"11","author":"Wei","year":"2019","journal-title":"Anal. Methods"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1016\/j.bios.2015.05.018","article-title":"Multi-color quantumdot-based fluorescence immunoassay array for simultaneous visual detection of multiple antibiotic residues in milk","volume":"72","author":"Song","year":"2015","journal-title":"Biosens. Bioelectron."},{"key":"ref_13","first-page":"374","article-title":"Rapid and sensitive chemiluminescent enzyme immunoassay for the determination of neomycin residues in milk","volume":"29","author":"Luo","year":"2016","journal-title":"Biomed. Environ. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1016\/j.cca.2005.07.024","article-title":"Development of ELISA and immunochromatographic assay for the detection of neomycin","volume":"364","author":"Jin","year":"2006","journal-title":"Clin. Chim. Acta"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"292","DOI":"10.1007\/s13197-020-04541-z","article-title":"Sensitive lateral flow immunoassay of an antibiotic neomycin","volume":"58","author":"Hendrickson","year":"2021","journal-title":"J. Food Sci. Technol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"9580","DOI":"10.1039\/C8RA01116H","article-title":"Development of an immunochromatographic lateral flow strip for the simultaneous detection of aminoglycoside residues in milk","volume":"8","author":"Sun","year":"2018","journal-title":"RSC Adv."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.aca.2020.06.009","article-title":"Using bimetallic Au@Pt nanozymes as a visual tag and as an enzyme mimic in enhanced sensitive lateral-flow immunoassays: Application for the detection of streptomycin","volume":"1126","author":"Wei","year":"2020","journal-title":"Anal. Chim. Acta"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"143129","DOI":"10.1016\/j.scitotenv.2020.143129","article-title":"Recent advances in aptamer-based sensors for aminoglycoside antibiotics detection and their applications","volume":"762","author":"Yue","year":"2021","journal-title":"Sci. Total Environ."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1016\/j.ab.2013.03.028","article-title":"A proof-of-concept receptor-based assay for sulfonamides","volume":"438","author":"Liang","year":"2013","journal-title":"Anal. Biochem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.bios.2015.03.016","article-title":"A highly sensitive and class-specific fluorescence polarisation assay for sulphonamides based on dihydropteroate synthase","volume":"70","author":"Wang","year":"2015","journal-title":"Biosens. Bioelectron."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.aca.2018.11.005","article-title":"Dihydropteroate synthase based sensor for screening multi-sulfonamides residue and its comparison with broad-specific antibody based immunoassay by molecular modeling analysis","volume":"1050","author":"Liang","year":"2019","journal-title":"Anal. Chim. Acta"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"13953","DOI":"10.1021\/acs.jafc.1c05674","article-title":"Development of a dihydropteroate synthase based fluorescence polarization assay for detection of sulfonamides and studying its recognition mechanism","volume":"69","author":"He","year":"2021","journal-title":"J. Agric. Food Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3023","DOI":"10.1021\/acs.jafc.2c00337","article-title":"Production of a natural dihydropteroate synthase and development of a signal amplified pseudo immunoassay for determination of sulfonamides in pork","volume":"70","author":"He","year":"2022","journal-title":"J. Agric. Food Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"104354","DOI":"10.1016\/j.microc.2019.104354","article-title":"Development and validation of an enzyme-linked receptor assay based on mutant protein I188K\/S19C\/G24C for 40 beta-lactams antibiotics detection in 13 food samples","volume":"152","author":"Ahmed","year":"2020","journal-title":"Microchem. J."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.ab.2018.10.017","article-title":"A receptor-based chemiluminescence enzyme linked immunosorbent assay for determination of tetracyclines in milk","volume":"564\u2013565","author":"Wang","year":"2019","journal-title":"Anal. Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"104184","DOI":"10.1016\/j.microc.2019.104184","article-title":"Directional evolution of TetR protein and development of a fluoroimmunoassay for screening of tetracyclines in egg","volume":"150","author":"Wang","year":"2019","journal-title":"Microchem. J."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"106779","DOI":"10.1016\/j.microc.2021.106779","article-title":"Synthesis of photoaffinity labeled activity-based protein profiling probe and production of natural TetR protein for immunoassay of tetracyclines in milk","volume":"170","author":"Xia","year":"2021","journal-title":"Microchem. J."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.aca.2007.02.057","article-title":"Solubilisation and binding characteristics of a recombinant beta2-adrenergic receptor expressed in the membrane of Escherichia coli for the multianalyte detection of beta-agonists and antagonists residues in food-producing animals","volume":"589","author":"Danyi","year":"2007","journal-title":"Anal. Chim. Acta"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1021\/cr0301088","article-title":"Molecular insights into aminoglycoside action and resistance","volume":"105","author":"Magnet","year":"2005","journal-title":"Chem. Rev."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"340","DOI":"10.1038\/35030019","article-title":"Functional insights from the structure of the 30S ribosomal subunit and its interactions with antibiotics","volume":"407","author":"Carter","year":"2000","journal-title":"Nature"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1075","DOI":"10.1039\/C4MD00163J","article-title":"New trends in the use of aminoglycosides","volume":"5","author":"Fosso","year":"2014","journal-title":"Med. Chem. Commun."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1128\/AAC.40.4.1024","article-title":"Characterization of rpsL and rrs mutations in streptomycin-resistant Mycobacterium tuberculosis isolates from diverse geographic localities","volume":"40","author":"Sreevatsan","year":"1996","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2080","DOI":"10.1261\/rna.302006","article-title":"A novel single amino acid change in small subunit ribosomal protein S5 has profound effects on translational fidelity","volume":"12","author":"Kirthi","year":"2006","journal-title":"RNA"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1128\/AAC.00388-08","article-title":"A novel insertion mutation in streptomyces coelicolorribosomal S12 protein results in paromomycin resistance and antibiotic overproduction","volume":"53","author":"Wang","year":"2009","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1190","DOI":"10.1080\/07391102.2015.1073633","article-title":"Streptomycin affinity depends on 13 amino acids forming a loop in homology modeled ribosomal S12 protein (rpsL gene) of Lysinibacillussphaericus DSLS5 associated with marine sponge (Tedaniaanhelans)","volume":"34","author":"Suriyanarayanan","year":"2016","journal-title":"J. Biomol. Struct. Dyn."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1016\/j.trac.2019.03.013","article-title":"Fluorescence polarization assays for chemical contaminants in food and environmental analyses","volume":"114","author":"Zhang","year":"2019","journal-title":"TRAC Trends Anal. Chem."},{"key":"ref_37","unstructured":"(2019). National Food Safety Standard\u2014Maximum Residue Limits for Veterinary Drugs in Foods (Standard No. GB 31650-2019)."}],"container-title":["Foods"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2304-8158\/11\/20\/3196\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:53:28Z","timestamp":1760144008000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2304-8158\/11\/20\/3196"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,13]]},"references-count":37,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2022,10]]}},"alternative-id":["foods11203196"],"URL":"https:\/\/doi.org\/10.3390\/foods11203196","relation":{},"ISSN":["2304-8158"],"issn-type":[{"value":"2304-8158","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,13]]}}}