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Canine seropositivity often correlates with human CD prevalence. However, the lack of commercial, high-performance diagnostic assays for canine infections remains a significant barrier to effective surveillance. Previously, our group demonstrated the diagnostic potential of four chimeric\n                      <jats:italic>T. cruzi<\/jats:italic>\n                      antigens in a phase I study, yielding results comparable to those observed in humans. The present phase II study expands upon these findings by evaluating these antigens in a larger canine cohort using indirect enzyme-linked immunosorbent assay (ELISA). The objective of this study was to assess the diagnostic performance of four chimeric recombinant\n                      <jats:italic>T. cruzi<\/jats:italic>\n                      antigens (IBMP-8.1, IBMP-8.2, IBMP-8.3, and IBMP-8.4) in immunoassays for the detection of anti-\n                      <jats:italic>T. cruzi<\/jats:italic>\n                      IgG in dogs with chronic Chagas disease.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Immunoassays were optimized by checkerboard titration. In this phase II study, the diagnostic performance of the IBMP antigens was evaluated using 1260 canine serum samples. Cross-reactivity was assessed in an additional 752 samples from dogs with unrelated infectious diseases. The performance of the chimeric antigens was compared with a commercial human-adapted assay (Gold ELISA Chagas).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The Instituto de Biologia Molecular do Paran\u00e1 (IBMP) antigens demonstrated area under the curve (AUC) values ranging from 89.0% to 97.4%, with diagnostic accuracy between 87.4% and 96%. IBMP-8.2 exhibited the highest sensitivity (90.3%), while IBMP-8.1, IBMP-8.3, and IBMP-8.4 achieved sensitivities of 74.8%, 72.6%, and 79.6%, respectively. The highest specificity was observed for IBMP-8.4 (99.6%), followed by IBMP-8.3 (99.0%), IBMP-8.2 (96.5%), and IBMP-8.1 (90.6%). The Gold ELISA Chagas assay showed a sensitivity of 62.3%, specificity of 98.6%, and accuracy of 89.9%. IBMP-8.2 exhibited the lowest cross-reactivity index (0.9%), closely approximating an ideal diagnostic assay.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      The IBMP chimeric antigens demonstrated strong diagnostic performance for detecting\n                      <jats:italic>T. cruzi<\/jats:italic>\n                      infection in dogs, significantly enhancing immunoassay accuracy and minimizing diagnostic failures due to cross-reactivity. The combined use of these antigens represents a promising strategy to further improve sensitivity and specificity in future diagnostic applications.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Graphical Abstract<\/jats:title>\n                  <\/jats:sec>","DOI":"10.1186\/s13071-025-07173-4","type":"journal-article","created":{"date-parts":[[2025,12,7]],"date-time":"2025-12-07T13:54:27Z","timestamp":1765115667000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Evaluation of four chimeric Trypanosoma cruzi recombinant antigens for serological diagnosis of chronic Chagas disease in dogs: a phase II study"],"prefix":"10.1186","volume":"19","author":[{"given":"Nat\u00e1lia Dantas","family":"Fontes","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernanda Lopes","family":"Habib","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leonardo Maia","family":"Leony","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalia Erdens Maron","family":"Freitas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c2ngelo Ant\u00f4nio Oliveira","family":"Silva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Filipe","family":"Dantas-Torres","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kamila Gaud\u00eancio da Silva","family":"Sales","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e9a Pereira da","family":"Costa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thaliane Fran\u00e7a","family":"Costa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nayara Mendes","family":"Louzeiro","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sidilene Pereira","family":"Costa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lileia Gon\u00e7alves","family":"Diotaiuti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlota Josefovcz","family":"Belisario","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cl\u00e1udia Moura de","family":"Melo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ant\u00f4nia Cl\u00e1udia J\u00e1come da","family":"C\u00e2mara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vicente Toscano de","family":"Ara\u00fajo-Neto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luanna Soares de Melo","family":"Evangelista","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deborah Bittencourt","family":"Moth\u00e9","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paola Alejandra Fiorani","family":"Celedon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nilson Ivo Tonin","family":"Zanchin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fred Luciano Neves","family":"Santos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,12,7]]},"reference":[{"key":"7173_CR1","volume-title":"Actualizaci\u00f3n de la Estimaci\u00f3n de la Enfermedad de Chagas en los Pa\u00edses End\u00e9micos de las Am\u00e9ricas 2018","author":"PAHO","year":"2025","unstructured":"PAHO. 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