ArticleBMC veterinary research2025
Development and application of ActA-based competitive ELISA for the specific diagnosis of ovine listeriosis.
Article in BMC veterinary research, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
backgroundListeriosis is a severe zoonotic disease caused by Listeria monocytogenes (L. monocytogenes), which poses a significant threat to both livestock and public health. Efficient detection of L. monocytogenes infections in humans and various animal species requires the development of a specific and sensitive method for clinical diagnosis.
resultsIn this study, a competitive enzyme-linked immunosorbent assay (cELISA) was developed for the detection of anti-ActA antibodies in serum, based on a horseradish peroxidase-labeled monoclonal antibody 3G11 (HRP-MAb 3G11). The assay parameters, including the coating antigen concentration, HRP-MAb 3G11 titer, serum dilution factor, blocking buffer, and blocking incubation time, were optimized for the detection of ActA-specific antibodies. Receiver operating characteristic (ROC) analysis identified 27.50% as the optimal cutoff value for the percentage of inhibition (PI), yielding a diagnostic sensitivity of 100% and a diagnostic specificity of 93.33%. This assay exhibited no cross-reactivity with positive sera specific for other common pathogens, demonstrated stable reproducibility, and achieved a diagnostic accuracy of 95.7% compared with a commercial indirect LLO-based ELISA kit. The cELISA kit was used to detect 930 ovine serum samples, revealing an overall seropositivity rate of 24.84%.
conclusionsThe cELISA kit developed in this study provides a specific, sensitive, and repeatable serological method for the rapid detection of L. monocytogenes infection.
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