| Author | Macedo, Priscila Marques de | |
| Author | Sturny-Leclère, Aude | |
| Author | Hamane, Samia | |
| Author | Pautet, Thierry | |
| Author | Rodrigues, Anderson Messias | |
| Author | Freitas, Dayvison Francis Saraiva | |
| Author | Valle, Antonio Carlos Francesconi do | |
| Author | Zancopé-Oliveira, Rosely Maria | |
| Author | Almeida-Paes, Rodrigo | |
| Author | Alanio, Alexandre | |
| Access date | 2025-03-13T18:51:40Z | |
| Available date | 2025-03-13T18:51:40Z | |
| Document date | 2025 | |
| Citation | MACEDO, Priscila Marques de et al. A new quantitative reverse transcription PCR assay to improve the routine diagnosis of paracoccidioidomycosis. Medical Mycology, v. 63, n. 1, p. 1-7, Jan. 2025. | en_US |
| ISSN | 1369-3786 | en_US |
| URI | https://www.arca.fiocruz.br/handle/icict/68971 | |
| Description | Lay summary: Paracoccidioidom y cosis (PCM) is a se v ere sy stemic m y cosis with limited laboratory techniques for rapid diagnosis. We de v eloped an assa y to detect genetic material of fungal agents of PCM and validated it in human specimens from patients with this disease, obtaining high positivity and specificity. | en_US |
| Sponsorship | The Instituto Nacional de Infectologia Evandro Chagas, Fundação Oswaldo Cruz (INI, Fiocruz), the Institut Pasteur, and the Hôpital Saint-Louis (Assistance Publique –Hôpitaux de Paris) supported this study. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brasil –Finance Code 001. This work was also supported in part by Fundação Carlos Chagas Filho de Amparo àPesquisa do Estado do Rio de Janeiro (FAPERJ), grant number E-26/211.430/2021 to PMdM. A.A. was funded by the French National Research Agency (ANR 20CE35000701). R.M.Z.-O was supported in part by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq 308315/2021-9) and (FAPERJ E-26/200.381/2023). A.M.R. was supported by the São Paulo Research Foundation (FAPESP 2017/27265-5) and is a CNPq Research Productivity Fellow (CNPq 314089/2023-3). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. | en_US |
| Language | eng | en_US |
| Publisher | Oxford | en_US |
| Rights | restricted access | en_US |
| Title | A new quantitative reverse transcription PCR assay to improve the routine diagnosis of paracoccidioidomycosis | en_US |
| Type | Article | |
| Abstract | Paracoccidioides are dimorphic fungal pathogens and the etiological agents of paracoccidioidomycosis (PCM). 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's specificity. In conclusion, this study proposes a new accurate tool to cover gaps, contributing to the molecular diagnosis of this neglected disease. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatologia Infecciosa. Rio de Janeiro, RJ, Brasil / Université Paris Cité. Institut Pasteur. Translational Mycology Research Group. National Reference Center for Invasive Mycoses and Antifungals. Mycology Department. Paris, France. | en_US |
| Affilliation | Université Paris Cité. Institut Pasteur. Translational Mycology Research Group. National Reference Center for Invasive Mycoses and Antifungals. Mycology Department. Paris, France. | en_US |
| Affilliation | Hôpital Saint-Louis. AP-HP. Laboratoire de Parasitologie-Mycologie. Paris, France. | en_US |
| Affilliation | Hôpital Saint-Louis. AP-HP. Laboratoire de Parasitologie-Mycologie. Paris, France. | en_US |
| Affilliation | Universidade Federal de São Paulo. Laboratório de Patógenos Fúngicos Emergentes. Departamento de Microbiologia, Imunologia e Parasitologia. São Paulo, SP, Brasil. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatologia Infecciosa. Rio de Janeiro, RJ, Brasil. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatologia Infecciosa. Rio de Janeiro, RJ, Brasil. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Micologia. Rio de Janeiro, RJ, Brasil. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Micologia. Rio de Janeiro, RJ, Brasil. | en_US |
| Affilliation | Université Paris Cité. Institut Pasteur. Translational Mycology Research Group. National Reference Center for Invasive Mycoses and Antifungals. Mycology Department. Paris, France / Hôpital Saint-Louis. AP-HP. Laboratoire de Parasitologie-Mycologie. Paris, France. | en_US |
| Subject | Paracoccidioidomycosis | en_US |
| Subject | Paracoccidioides | en_US |
| Subject | Endemic mycosis | en_US |
| Subject | Molecular diagnosis | en_US |
| Embargo date | 2030-12-31 | |
