Development of an internalin-based double-antibody sandwich quantitative ELISA for the detection of Listeria monocytogenes in slaughterhouse environments.
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Abstract
Introduction: Listeria monocytogenes causes zoonotic listeriosis with a high mortality rate, which is frequently detected in slaughterhouse processing environments and animal-based food. To enable the specific, rapid, and cost-effective detection of L. monocytogenes in environments and animal-based food, we developed a double-antibody sandwich quantitative ELISA (DAS-qELISA) method.
Methods: The method is based on monoclonal antibodies targeting internalin G (InlG), a surface protein of L. monocytogenes with demonstrated immunogenicity. The antibody pair 1D2-2H10 was selected for use in the sandwich ELISA format. Optimization of the DAS-qELISA method was carried out to determine its detection limits for InlG protein and L. monocytogenes.
Results: The detection limits of the method were determined to be 32 ng/mg for the InlG protein and 7875.83 CFU/mL for L. monocytogenes. The accuracy of the method was evaluated across various bacterial concentrations, with results falling within 91.56-107.07% and a coefficient of variation (CV) of less than 10%. Compared to traditional methods, this approach requires only 12 h of bacterial enrichment and incubation to achieve 100% accuracy.
Discussion: The DAS-qELISA developed in this study provides a rapid, accurate, and cost-effective tool for the detection of L. monocytogenes in environmental and animal-based food samples. This method could be a valuable addition to current diagnostic approaches, offering quicker turnaround times and high accuracy for pathogen detection.
期刊介绍：
Frontiers in Veterinary Science is a global, peer-reviewed, Open Access journal that bridges animal and human health, brings a comparative approach to medical and surgical challenges, and advances innovative biotechnology and therapy.
Veterinary research today is interdisciplinary, collaborative, and socially relevant, transforming how we understand and investigate animal health and disease. Fundamental research in emerging infectious diseases, predictive genomics, stem cell therapy, and translational modelling is grounded within the integrative social context of public and environmental health, wildlife conservation, novel biomarkers, societal well-being, and cutting-edge clinical practice and specialization. Frontiers in Veterinary Science brings a 21st-century approach—networked, collaborative, and Open Access—to communicate this progress and innovation to both the specialist and to the wider audience of readers in the field.
Frontiers in Veterinary Science publishes articles on outstanding discoveries across a wide spectrum of translational, foundational, and clinical research. The journal''s mission is to bring all relevant veterinary sciences together on a single platform with the goal of improving animal and human health.
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