ORIGINAL RESEARCH article
Front. Vet. Sci.
Sec. Veterinary Clinical, Anatomical, and Comparative Pathology
Volume 12 - 2025 | doi: 10.3389/fvets.2025.1636223
This article is part of the Research TopicGo with the Vet-Flow! The Current Uses and New Frontiers of Flow Cytometry in Veterinary Sciences - Volume IIView all 5 articles
Rapid Detection of Antimicrobial Residues in Broiler Meat Using Flow Cytometry-Driven Multiplex Immunoassay
Provisionally accepted- 1Department of Veterinary Science, University of Turin, Grugliasco, Italy
- 2Department of Veterinary Medicine and Animal Science, University of Milan, Lodi, Italy
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The misuse of antimicrobials (AMs) in poultry production contributes significantly to the global threat of antimicrobial resistance. Despite legislative efforts within the European Union (EU) to reduce AM use, Italy remains among the countries with the highest consumption levels. Effective and accessible screening tools for AMs residues in poultry meat are crucial for food safety monitoring and regulatory compliance. This study evaluates the performance of the Beadyplex flow cytometric assay as a field-applicable method to detect residues of thiamphenicol, sulphadiazine, and amoxicillin in broiler chicken skeletal muscle. Pectoralis major muscle samples were collected from 41 male broiler chickens (Ross 308) previously treated with thiamphenicol (n=11), sulphadiazine (n=6), amoxicillin (n=12), or untreated (controls, n=12). Beadyplex assays were performed following manufacturer instructions, and results were acquired using a standard flow cytometer. Sensitivity, specificity, predictive values, and kappa agreement were calculated to assess the performance of the test in recognizing the AMs treatment. The agreement with the current reference method for detecting AM residues (high-performance liquid chromatography–high-resolution mass spectrometry, or HPLC-HRMS) was also calculated. The Beadyplex assay demonstrated high sensitivity and specificity for thiamphenicol (Se = 1.00; Sp = 0.83; k = 0.791) and sulphadiazine (Se = 1.00; Sp = 0.91; k = 0.813), with substantial to almost perfect agreement with HPLC-HRMS results (k = 0.706 and 0.827, respectively). Amoxicillin residues were not detected in any treated sample by Beadyplex. The Beadyplex assay offers a promising, cost-effective, and rapid screening complementary to confirmatory analysis for detecting selected AM residues in poultry meat. Its substantial agreement with reference methods and ease of use with standard cytometers support its potential application in field-based veterinary surveillance and food safety programs.
Keywords: Beadyplex, antimicrobials, Immunoassay, Flow Cytometry, HPLC-HRMS
Received: 27 May 2025; Accepted: 29 Sep 2025.
Copyright: © 2025 Cannizzo, Sini, Divari, Cuccato, Poggi, Panseri, Nobile, Chiesa and RIONDATO. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Matteo Cuccato, matteo.cuccato@unito.it
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