{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T08:06:50Z","timestamp":1789546010389,"version":"build-2803163510"},"reference-count":36,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T00:00:00Z","timestamp":1789516800000},"content-version":"vor","delay-in-days":258,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Veterinary Medicine International"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:p>\n                    <jats:italic>Gallibacterium anatis<\/jats:italic>\n                    is an emerging opportunistic pathogen associated with respiratory and reproductive tract infections in poultry, causing reduced egg production and economic losses in laying hens. Serological detection is important for surveillance; however, no validated in\u2010house enzyme\u2010linked immunosorbent assay (ELISA) based on local Indonesian isolates is available. This study aimed to develop and validate an indirect ELISA for detecting antibodies against\n                    <jats:italic>G. anatis<\/jats:italic>\n                    in laying hens. Five archived isolates were characterized by PCR targeting gyrB, gtxA, and flfA virulence\u2010associated genes, followed by growth rate and biofilm formation analyses. Isolate B116, which showed the highest growth rate and strongest biofilm\u2010forming ability, was selected for antigen preparation. Four antigen extraction methods, including heat treatment at 56\u00b0C and 96\u00b0C, capsular extraction, and sonication, were evaluated to determine the best coating antigen. Checkerboard titration was performed to optimize antigen concentration, serum dilution, and HRP\u2010conjugated rabbit anti\u2010chicken IgG dilution. Cross\u2010reactivity was assessed using positive sera against\n                    <jats:italic>Pasteurella multocida<\/jats:italic>\n                    ,\n                    <jats:italic>Avibacterium paragallinarum, Escherichia coli<\/jats:italic>\n                    ,\n                    <jats:italic>Salmonella sp.<\/jats:italic>\n                    , and\n                    <jats:italic>Clostridium perfringens<\/jats:italic>\n                    . The optimal ELISA conditions were 7.11\u2009\u03bcg\/mL coating antigen concentration, 1:200 serum dilution, and 1:8000 conjugate dilution. The cutoff values were 0.156 and 0.189 using mean \u00b1 2SD and mean \u00b1 3SD. Compared with PCR, the in\u2010house ELISA showed 85.0% sensitivity and 94.7% specificity. Field serum testing revealed the highest seropositivity in laying hens aged 29\u201348\u2009weeks. The developed in\u2010house ELISA demonstrated good diagnostic performance and may serve as a practical tool for large\u2010scale serological surveillance and epidemiological monitoring of\n                    <jats:italic>G. anatis<\/jats:italic>\n                    infection in poultry farms.\n                  <\/jats:p>","DOI":"10.1155\/vmi\/9467618","type":"journal-article","created":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T07:43:03Z","timestamp":1789544583000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and Validation of an In\u2010House Indirect ELISA for Serological Detection of\n                    <i>Gallibacterium anatis<\/i>\n                    in Laying Hens Using Local Field Isolates"],"prefix":"10.1155","volume":"2026","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-3307-0589","authenticated-orcid":false,"given":"Raudatul","family":"Jannah","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3135-009X","authenticated-orcid":false,"given":"Agustin","family":"Indrawati","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9554-3902","authenticated-orcid":false,"given":"Ryan Septa","family":"Kurnia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1940-2777","authenticated-orcid":false,"given":"Christian Marco Hadi","family":"Nugroho","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2828-1399","authenticated-orcid":false,"given":"Muhammad Ade","family":"Putra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6251-8735","authenticated-orcid":false,"given":"Sri","family":"Murtini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,16]]},"reference":[{"key":"e_1_2_11_1_2","first-page":"1301","article-title":"Dot-ELISA for Newcastle Disease","volume":"44","author":"Alam J.","year":"2012","journal-title":"Infectious Bursal Disease and Mycoplasmosis"},{"key":"e_1_2_11_2_2","doi-asserted-by":"publisher","DOI":"10.1051\/vetres:2001125"},{"key":"e_1_2_11_3_2","doi-asserted-by":"publisher","DOI":"10.1080\/01652176.2020.1712495"},{"key":"e_1_2_11_4_2","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms8101533"},{"key":"e_1_2_11_5_2","doi-asserted-by":"publisher","DOI":"10.54203\/scil.2024.wvj2"},{"key":"e_1_2_11_6_2","doi-asserted-by":"publisher","DOI":"10.21608\/avmj.2015.170239"},{"key":"e_1_2_11_7_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13567-021-00894-1"},{"key":"e_1_2_11_8_2","doi-asserted-by":"publisher","DOI":"10.5808\/gi.22006"},{"key":"e_1_2_11_9_2","doi-asserted-by":"publisher","DOI":"10.4236\/aim.2012.24064"},{"key":"e_1_2_11_10_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2023.1084766"},{"key":"e_1_2_11_11_2","doi-asserted-by":"publisher","DOI":"10.21474\/ijar01\/8226"},{"key":"e_1_2_11_12_2","doi-asserted-by":"publisher","DOI":"10.1080\/03079457.2013.843160"},{"key":"e_1_2_11_13_2","unstructured":"ZhangJ. 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