{"id":27625,"date":"2025-09-16T12:31:26","date_gmt":"2025-09-16T04:31:26","guid":{"rendered":"https:\/\/tbenews.com\/tbe\/?p=27625"},"modified":"2025-09-16T12:31:48","modified_gmt":"2025-09-16T04:31:48","slug":"snapshot-week-39-2025-mutant-e-protein-elisa-for-differentiating-flavivirus-antibodies-in-birds-and-horses","status":"publish","type":"post","link":"https:\/\/tbenews.com\/tbe\/snapshot-week-39-2025-mutant-e-protein-elisa-for-differentiating-flavivirus-antibodies-in-birds-and-horses\/","title":{"rendered":"Snapshot week 39\/2025<br>Mutant E protein ELISA for differentiating flavivirus antibodies in birds and horses"},"content":{"rendered":"<span class=\"cb-itemprop\" itemprop=\"reviewBody\">\n<p class=\"wp-block-paragraph\">Schwarzer et al.<br>Serological differentiation of West Nile, Usutu, and tick-borne encephalitis virus antibodies in birds and horses using mutant E protein ELISAs. <em>Scientific Reports<\/em>. 2025;15:28752. doi:10.1038\/s41598-025-14448-4.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">Specific serological diagnosis of flavivirus infections is challenging due to the strong antigenic similarity among closely related flaviviruses. This similarity leads to highly cross-reactive antibody responses, particularly in ELISA systems based on the flaviviral envelope glycoprotein E.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">To address this, a novel ELISA has been developed in which specific mutations were introduced into the fusion loop (FL) of the envelope glycoprotein E (gE). These mutations enable reliable differentiation of West Nile virus, Usutu virus, and tick-borne encephalitis (TBE) virus infections in horses and birds. For assay development, the ectodomains of gE for each of the three viruses were engineered to contain four-point mutations near the FL domain and were expressed in a Drosophila S2 cell system.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">This newly established ELISA has the potential to replace the virus neutralization test, which is currently considered the gold standard but requires biosafety level 3 facilities. Using the modified ELISA, the distribution of co-circulating flaviviruses in parts of Germany was analyzed in both avian (ducks, geese, chickens) and equine sera.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">By incorporating a modified FL domain, the assay effectively minimized cross-reactive binding among closely related West Nile, Usutu, and TBE virus IgY and IgG antibodies. The test system demonstrated high specificity and sensitivity, while being rapid, cost-effective, and sustainable. Importantly, it allows for reliable and efficient serological diagnosis of flavivirus infections in standard diagnostic laboratories and for use in seroprevalence studies.<\/p>\n<\/span>","protected":false},"excerpt":{"rendered":"<p>Schwarzer et al.Serological differentiation of West Nile, Usutu, and tick-borne encephalitis virus antibodies in birds and horses using mutant E protein ELISAs. Scientific Reports. 2025;15:28752. doi:10.1038\/s41598-025-14448-4. Specific serological diagnosis of flavivirus infections is challenging due to the strong antigenic similarity among closely related flaviviruses. This similarity leads to highly cross-reactive antibody responses, particularly in ELISA systems based on the flaviviral envelope glycoprotein E. To address this, a novel ELISA has been developed in which specific mutations were introduced into the fusion loop (FL) of the envelope glycoprotein E (gE). These mutations enable reliable differentiation of West Nile virus, Usutu virus, and tick-borne encephalitis (TBE) virus infections in horses and birds. For assay development, the ectodomains of gE for each of the three viruses were engineered to contain four-point mutations near the FL domain and were expressed in a Drosophila S2 cell system. This newly established ELISA has the potential to replace the virus neutralization test, which is currently considered the gold standard but requires biosafety level 3 facilities. Using the modified ELISA, the distribution of co-circulating flaviviruses in parts of Germany was analyzed in both avian (ducks, geese, chickens) and equine sera. By incorporating a modified FL domain, the assay effectively minimized cross-reactive binding among closely related West Nile, Usutu, and TBE virus IgY and IgG antibodies. The test system demonstrated high specificity and sensitivity, while being rapid, cost-effective, and sustainable. Importantly, it allows for reliable and efficient serological diagnosis of flavivirus infections in standard diagnostic laboratories and for use in seroprevalence studies.<\/p>\n","protected":false},"author":94681,"featured_media":20968,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_monsterinsights_skip_tracking":false,"footnotes":""},"categories":[18],"tags":[223,214,1295,264,781,60],"coauthors":[1108],"class_list":["post-27625","post","type-post","status-publish","format-standard","has-post-thumbnail","category-snapshot","tag-cross-reactivity","tag-elisa","tag-fusion-loop-domain","tag-glycoprotein-e","tag-usutu-virus","tag-west-nile-virus"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v28.1 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Snapshot week 39\/2025Mutant E protein ELISA for differentiating flavivirus antibodies in birds and horses - TBE Book<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/tbenews.com\/tbe\/snapshot-week-39-2025-mutant-e-protein-elisa-for-differentiating-flavivirus-antibodies-in-birds-and-horses\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Snapshot week 39\/2025Mutant E protein ELISA for differentiating flavivirus antibodies in birds and horses - TBE Book\" \/>\n<meta property=\"og:description\" content=\"Schwarzer et al.Serological differentiation of West Nile, Usutu, and tick-borne encephalitis virus antibodies in birds and horses using mutant E protein ELISAs. Scientific Reports. 2025;15:28752. doi:10.1038\/s41598-025-14448-4. Specific serological diagnosis of flavivirus infections is challenging due to the strong antigenic similarity among closely related flaviviruses. This similarity leads to highly cross-reactive antibody responses, particularly in ELISA systems based on the flaviviral envelope glycoprotein E. To address this, a novel ELISA has been developed in which specific mutations were introduced into the fusion loop (FL) of the envelope glycoprotein E (gE). These mutations enable reliable differentiation of West Nile virus, Usutu virus, and tick-borne encephalitis (TBE) virus infections in horses and birds. For assay development, the ectodomains of gE for each of the three viruses were engineered to contain four-point mutations near the FL domain and were expressed in a Drosophila S2 cell system. This newly established ELISA has the potential to replace the virus neutralization test, which is currently considered the gold standard but requires biosafety level 3 facilities. Using the modified ELISA, the distribution of co-circulating flaviviruses in parts of Germany was analyzed in both avian (ducks, geese, chickens) and equine sera. By incorporating a modified FL domain, the assay effectively minimized cross-reactive binding among closely related West Nile, Usutu, and TBE virus IgY and IgG antibodies. The test system demonstrated high specificity and sensitivity, while being rapid, cost-effective, and sustainable. Importantly, it allows for reliable and efficient serological diagnosis of flavivirus infections in standard diagnostic laboratories and for use in seroprevalence studies.\" \/>\n<meta property=\"og:url\" content=\"https:\/\/tbenews.com\/tbe\/snapshot-week-39-2025-mutant-e-protein-elisa-for-differentiating-flavivirus-antibodies-in-birds-and-horses\/\" \/>\n<meta property=\"og:site_name\" content=\"TBE Book\" \/>\n<meta property=\"article:published_time\" content=\"2025-09-16T04:31:26+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2025-09-16T04:31:48+00:00\" \/>\n<meta property=\"og:image\" content=\"https:\/\/tbenews.com\/tbe\/wp-content\/uploads\/2023\/12\/TBE-Snapshots.jpg\" \/>\n\t<meta property=\"og:image:width\" content=\"1180\" \/>\n\t<meta property=\"og:image:height\" content=\"250\" \/>\n\t<meta property=\"og:image:type\" content=\"image\/jpeg\" \/>\n<meta name=\"author\" content=\"logesan\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:label1\" content=\"Written by\" \/>\n\t<meta name=\"twitter:data1\" content=\"logesan\" \/>\n\t<meta name=\"twitter:label2\" content=\"Est. reading time\" \/>\n\t<meta name=\"twitter:data2\" content=\"2 minutes\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\\\/\\\/schema.org\",\"@graph\":[{\"@type\":\"Article\",\"@id\":\"https:\\\/\\\/tbenews.com\\\/tbe\\\/snapshot-week-39-2025-mutant-e-protein-elisa-for-differentiating-flavivirus-antibodies-in-birds-and-horses\\\/#article\",\"isPartOf\":{\"@id\":\"https:\\\/\\\/tbenews.com\\\/tbe\\\/snapshot-week-39-2025-mutant-e-protein-elisa-for-differentiating-flavivirus-antibodies-in-birds-and-horses\\\/\"},\"author\":{\"name\":\"logesan\",\"@id\":\"https:\\\/\\\/tbenews.com\\\/tbe\\\/#\\\/schema\\\/person\\\/eea00490409f8d848a28485d0f68fac3\"},\"headline\":\"Snapshot week 39\\\/2025 Mutant E protein ELISA for differentiating flavivirus antibodies in birds and horses\",\"datePublished\":\"2025-09-16T04:31:26+00:00\",\"dateModified\":\"2025-09-16T04:31:48+00:00\",\"mainEntityOfPage\":{\"@id\":\"https:\\\/\\\/tbenews.com\\\/tbe\\\/snapshot-week-39-2025-mutant-e-protein-elisa-for-differentiating-flavivirus-antibodies-in-birds-and-horses\\\/\"},\"wordCount\":270,\"publisher\":{\"@id\":\"https:\\\/\\\/tbenews.com\\\/tbe\\\/#organization\"},\"image\":{\"@id\":\"https:\\\/\\\/tbenews.com\\\/tbe\\\/snapshot-week-39-2025-mutant-e-protein-elisa-for-differentiating-flavivirus-antibodies-in-birds-and-horses\\\/#primaryimage\"},\"thumbnailUrl\":\"https:\\\/\\\/tbenews.com\\\/tbe\\\/wp-content\\\/uploads\\\/2023\\\/12\\\/TBE-Snapshots.jpg\",\"keywords\":[\"cross-reactivity\",\"ELISA\",\"fusion loop domain\",\"glycoprotein E\",\"Usutu virus\",\"West Nile virus\"],\"articleSection\":[\"Snapshot\"],\"inLanguage\":\"en-US\"},{\"@type\":\"WebPage\",\"@id\":\"https:\\\/\\\/tbenews.com\\\/tbe\\\/snapshot-week-39-2025-mutant-e-protein-elisa-for-differentiating-flavivirus-antibodies-in-birds-and-horses\\\/\",\"url\":\"https:\\\/\\\/tbenews.com\\\/tbe\\\/snapshot-week-39-2025-mutant-e-protein-elisa-for-differentiating-flavivirus-antibodies-in-birds-and-horses\\\/\",\"name\":\"Snapshot week 39\\\/2025 Mutant E protein ELISA for differentiating flavivirus antibodies in birds and horses - 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Scientific Reports. 2025;15:28752. doi:10.1038\/s41598-025-14448-4. Specific serological diagnosis of flavivirus infections is challenging due to the strong antigenic similarity among closely related flaviviruses. This similarity leads to highly cross-reactive antibody responses, particularly in ELISA systems based on the flaviviral envelope glycoprotein E. To address this, a novel ELISA has been developed in which specific mutations were introduced into the fusion loop (FL) of the envelope glycoprotein E (gE). These mutations enable reliable differentiation of West Nile virus, Usutu virus, and tick-borne encephalitis (TBE) virus infections in horses and birds. For assay development, the ectodomains of gE for each of the three viruses were engineered to contain four-point mutations near the FL domain and were expressed in a Drosophila S2 cell system. This newly established ELISA has the potential to replace the virus neutralization test, which is currently considered the gold standard but requires biosafety level 3 facilities. Using the modified ELISA, the distribution of co-circulating flaviviruses in parts of Germany was analyzed in both avian (ducks, geese, chickens) and equine sera. By incorporating a modified FL domain, the assay effectively minimized cross-reactive binding among closely related West Nile, Usutu, and TBE virus IgY and IgG antibodies. The test system demonstrated high specificity and sensitivity, while being rapid, cost-effective, and sustainable. 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