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This study pioneered integrating nanotechnology into an indirect enzyme-linked immunosorbent assay (i-ELISA) system to enhance the sensitivity and specificity for detecting <jats:italic>S. vulgaris<\/jats:italic> larval antigens in equine serum samples, with PCR confirmation of the species. A conventional i-ELISA and an innovative nano-based ELISA were developed using excretory-secretory antigens from adult <jats:italic>S. vulgaris<\/jats:italic> worms. The nano-ELISA incorporated gold nanoparticles (17.4\u201341.4\u00a0nm) conjugated with detection antibodies, enabling remarkable signal amplification. Of the 120 examined equines, 100 (83.33%) were positive for <jats:italic>S. vulgaris<\/jats:italic> infection. A conventional i-ELISA and an innovative nano-ELISA incorporating 17.4\u201341.4\u00a0nm gold nanoparticles were optimized using <jats:italic>S. vulgaris<\/jats:italic> excretory-secretory antigens. Both assays demonstrated high specificity, with no cross-reactivity against sera from animals infected with other helminth parasites. Remarkably, optical density (OD) readings from both i-ELISAs exhibited a positive quantitative correlation with infection intensity. The i-ELISA OD ranged from 0.45\u20130.74 (G3), 0.75\u20130.94 (G2), to 0.95\u20132.5 (G1). The nano-ELISA showed enhanced signal amplification, with OD ranging from 0.40\u20130.84 (G3), 0.85\u20130.99 (G2), to 1.0\u20133.5 (G1). This nanotechnology-amplified ELISA opens new, highly sensitive, and specific techniques for parasitic diagnosis in equine medicine. Its superior performance, facilitated by signal-amplifying gold nanoparticles, illuminates nanotechnology's potential in revolutionizing parasitological diagnostics for enhanced animal health and welfare management.<\/jats:p>","DOI":"10.1186\/s12917-024-04389-x","type":"journal-article","created":{"date-parts":[[2024,12,27]],"date-time":"2024-12-27T07:15:10Z","timestamp":1735283710000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Diagnostic innovations in Equine Parasitology: a Nanogold-ELISA for sensitive serodiagnosis of migratory strongylus vulgaris larvae infections"],"prefix":"10.1186","volume":"20","author":[{"given":"Hanadi B. 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All the methods were carried out in accordance with the relevant guidelines and regulations of the use and handling of the animals and we take a permission from our university for sampling.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"579"}}