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This severe systemic mycosis is restricted to Latin America, where it has been historically endemic. Currently, PCM presents the fewest diagnostic tools available when compared to other endemic mycoses. The main PCM diagnostic methods also have limitations. Molecular methods using different protocols have been proposed, but are restricted to a few regions. An analytical transversal study was conducted to evaluate a new molecular tool using specimens from patients diagnosed with PCM at a reference center for endemic mycoses in Rio de Janeiro, Brazil. After whole nucleic acid (WNA) extraction, RT-qPCR was performed in two independent simplex reactions, targeting the mitochondrial small subunit ribosomal RNA genes of Paracoccidioides brasiliensis and Paracoccidioides lutzii. Additionally, WNAs from all PCM-related Paracoccidioides species and from 114 other fungal strains, as well as from samples obtained from patients diagnosed with other endemic mycoses and tuberculosis, were also tested for specificity. The RT-qPCR targeting P. brasiliensis successfully amplified genetic material from all tested Paracoccidioides species but not P. lutzii, which is why a specific RT-qPCR was designed. The RT-qPCR efficiency was 1.95 (95%) with 100% analytical specificity for both targets. All included PCM clinical samples were positive (100% sensitivity) for P. brasiliensis, and all yielded negative for P. lutzii. Additionally, all samples collected from patients with other diseases were negative, reinforcing the assay\u2019s specificity. In conclusion, this study proposes a new accurate tool to cover gaps, contributing to the molecular diagnosis of this neglected disease.<\/jats:p>","DOI":"10.1093\/mmy\/myae125","type":"journal-article","created":{"date-parts":[[2025,1,9]],"date-time":"2025-01-09T02:28:42Z","timestamp":1736389722000},"source":"Crossref","is-referenced-by-count":2,"title":["A new quantitative reverse transcription PCR assay to improve the routine diagnosis of paracoccidioidomycosis"],"prefix":"10.1093","volume":"63","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6900-9703","authenticated-orcid":false,"given":"Priscila","family":"Marques\u00a0de\u00a0Macedo","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio de Pesquisa Cl\u00ednica em Dermatologia Infecciosa, Instituto Nacional de Infectologia Evandro Chagas (INI), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz) , Rio de Janeiro ,","place":["Brazil"]},{"name":"Mycology Department, National Reference Center for Invasive Mycoses and Antifungals, Translational Mycology Research Group, Institut Pasteur, Universit\u00e9 Paris Cit\u00e9 , Paris ,","place":["France"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aude","family":"Sturny-Lecl\u00e8re","sequence":"additional","affiliation":[{"name":"Mycology Department, National Reference Center for Invasive Mycoses and Antifungals, Translational Mycology Research Group, Institut Pasteur, Universit\u00e9 Paris Cit\u00e9 , Paris ,","place":["France"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Samia","family":"Hamane","sequence":"additional","affiliation":[{"name":"Laboratoire de Parasitologie-Mycologie, AP-HP, H\u00f4pital Saint-Louis , Paris ,","place":["France"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thierry","family":"Pautet","sequence":"additional","affiliation":[{"name":"Laboratoire de Parasitologie-Mycologie, AP-HP, H\u00f4pital Saint-Louis , Paris ,","place":["France"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anderson Messias","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Departamento de Microbiologia, Imunologia e Parasitologia, Laborat\u00f3rio de Pat\u00f3genos F\u00fangicos Emergentes, Universidade Federal de S\u00e3o Paulo (UNIFESP) , S\u00e3o Paulo ,","place":["Brazil"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dayvison Francis Saraiva","family":"Freitas","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Pesquisa Cl\u00ednica em Dermatologia Infecciosa, Instituto Nacional de Infectologia Evandro Chagas (INI), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz) , Rio de Janeiro ,","place":["Brazil"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonio Carlos Francesconi do","family":"Valle","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Pesquisa Cl\u00ednica em Dermatologia Infecciosa, Instituto Nacional de Infectologia Evandro Chagas (INI), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz) , Rio de Janeiro ,","place":["Brazil"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rosely Maria","family":"Zancop\u00e9-Oliveira","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Micologia, Instituto Nacional de Infectologia Evandro Chagas (INI), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz) , Rio de Janeiro ,","place":["Brazil"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6639-6058","authenticated-orcid":false,"given":"Rodrigo","family":"Almeida-Paes","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Micologia, Instituto Nacional de Infectologia Evandro Chagas (INI), Funda\u00e7\u00e3o Oswaldo Cruz (Fiocruz) , Rio de Janeiro ,","place":["Brazil"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9726-3082","authenticated-orcid":false,"given":"Alexandre","family":"Alanio","sequence":"additional","affiliation":[{"name":"Mycology Department, National Reference Center for Invasive Mycoses and Antifungals, Translational Mycology Research Group, Institut Pasteur, Universit\u00e9 Paris Cit\u00e9 , Paris ,","place":["France"]},{"name":"Laboratoire de Parasitologie-Mycologie, AP-HP, H\u00f4pital Saint-Louis , Paris ,","place":["France"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2025,1,8]]},"reference":[{"key":"2025021015301828900_bib1","first-page":"121","article-title":"Uma mycose pseudococcidica localisada na bocca e observada no Brazil. 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