Development of an internalin-based double-antibody sandwich quantitative ELISA for the detection of Listeria monocytogenes in slaughterhouse environments

被引:0
|
作者
Cao, Qing [1 ]
Shi, Wenjing [1 ]
Wei, Yanquan [1 ]
Wang, Jiayu [1 ]
Wang, Zhonglong [1 ]
Chong, Qian [1 ]
Guo, Qianqian [1 ]
Zhang, Kunzhong [1 ]
Gai, Wenyan [2 ]
Gou, Huitian [1 ]
Xue, Huiwen [1 ]
机构
[1] Gansu Agr Univ, Coll Vet Med, Lanzhou, Peoples R China
[2] Shandong Vocat Anim Sci & Vet Coll, Weifang, Peoples R China
关键词
Listeria monocytogenes; internalin G; monoclonal antibodies; slaughterhouse; DAS-qELISA method; PREVALENCE; PROTEINS; BINDING; FOOD; PCR;
D O I
10.3389/fvets.2025.1517845
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
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.
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页数:10
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