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The existing methods for Paracoccidioides spp. antigen production are problematic in terms of standardization, specificity, stability, repeatability, and reproducibility.Objective. To optimize the methodology for Paracoccidioides spp. antigen production and evaluate its applicability in paracoccidioidomycosis immunodiagnosis.Materials and methods. The antigens were obtained from Paracoccidioides lutzii isolates (01, 66, and 8334), Paracoccidioides brasiliensis sensu stricto (113), and Paracoccidioides restripiensis (B-339). These fungi were grown at 36 \u00b0C \u00b1 1 \u00b0C, on modified Fava-Netto agar, according to Freitas et al. (2018). Paracoccidioides lutzii antigens were obtained after , 10, and 20 days of culture, whereas P. brasiliensis and P. restripiensis antigens were obtained after 10 days. Antigens were evaluated in natura, 10 and 20 times concentrated. Antigenic capacity was evaluated using a double immunodiffusion assay against serum samples from patients with paracoccidioidomycosis, histoplasmosis, and aspergillosis, and random blood donors.Results. Cross-reactivity between Paracoccidioides spp. antigens was observed when P. brasiliensis, P. restrepiensis antigens, and P. lutzii antigens were evaluated with the polyclonal antibodies against P. lutzii and P. brasiliensis, respectively. No cross-reactivity was obtained for polyclonal antibodies against Histoplasma capsulatum, Aspergillus fumigatus, and random blood donors. The proposed protocol allowed stable, repeatable, and reproducible genus-specific antigen production at a low cost and in a short cultivation time.Conclusion. The proposed protocol allowed us to obtain genus-specific antigens that can be developed and reproduced in all laboratories in Brazil and South America, where paracoccidioidomycosis is a neglected disease, contributing to an early diagnosis, especially in endemic regions, regardless of the species.<\/jats:p>","DOI":"10.7705\/biomedica.6874","type":"journal-article","created":{"date-parts":[[2023,9,18]],"date-time":"2023-09-18T10:18:35Z","timestamp":1695032315000},"page":"170-180","update-policy":"https:\/\/doi.org\/10.7705\/crossmark","source":"Crossref","is-referenced-by-count":2,"title":["Fast and cost-effective protocol to produce Paracoccidioides spp. antigens","Protocolo r\u00e1pido y econ\u00f3mico para la producci\u00f3n de ant\u00edgenos de Paracoccidioides spp."],"prefix":"10.7705","volume":"43","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0462-5254","authenticated-orcid":false,"given":"Karolina Rosa","family":"Fernandes-Beraldo","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio Central, Universidade Federal de S\u00e3o Paulo, S\u00e3o Paulo, Brasil;\u00a0Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias, Coordenadoria de Controle de Doen\u00e7as, Secretaria de Estado da Sa\u00fade de S\u00e3o Paulo, S\u00e3o Paulo, Brasil"},{"name":"Laborat\u00f3rio Central, Universidade Federal de S\u00e3o Paulo, S\u00e3o Paulo, Brasil;\u00a0Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias, Coordenadoria de Controle de Doen\u00e7as, Secretaria de Estado da Sa\u00fade de S\u00e3o Paulo, S\u00e3o Paulo, Brasil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5811-5255","authenticated-orcid":false,"given":"Roseli","family":"Santos de Freitas-Xavier","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Micologia M\u00e9dica (LIM53), Hospital das Cl\u00ednicas, Faculdade de Medicina, Universidade de S\u00e3o Paulo, S\u00e3o Paulo, Brasil"},{"name":"Laborat\u00f3rio de Micologia M\u00e9dica (LIM53), Hospital das Cl\u00ednicas, Faculdade de Medicina, Universidade de S\u00e3o Paulo, S\u00e3o Paulo, Brasil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9321-0909","authenticated-orcid":false,"given":"Adriana","family":"Pardini-Vicentini","sequence":"additional","affiliation":[{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias, Coordenadoria de Controle de Doen\u00e7as, Secretaria de Estado da Sa\u00fade de S\u00e3o Paulo, S\u00e3o Paulo, Brasil; Laborat\u00f3rio de Imunodiagn\u00f3stico das Micoses, Centro de Imunologia, Instituto Adolfo Lutz, S\u00e3o Universidade de S\u00e3o Paulo, S\u00e3o Paulo, Brasil"},{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias, Coordenadoria de Controle de Doen\u00e7as, Secretaria de Estado da Sa\u00fade de S\u00e3o Paulo, S\u00e3o Paulo, Brasil; Laborat\u00f3rio de Imunodiagn\u00f3stico das Micoses, Centro de Imunologia, Instituto Adolfo Lutz, S\u00e3o Universidade de S\u00e3o Paulo, S\u00e3o Paulo, Brasil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"4444","published-online":{"date-parts":[[2023,8,31]]},"reference":[{"key":"124060","doi-asserted-by":"crossref","unstructured":"Gonzalez A, Hernandez O. 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