ROSELI SANTOS DE FREITAS

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LIM/53 - Laboratório de Micologia, Hospital das Clínicas, Faculdade de Medicina
LIM/05 - Laboratório de Poluição Atmosférica Experimental, Hospital das Clínicas, Faculdade de Medicina

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  • article 0 Citação(ões) na Scopus
    Fast and cost-effective protocol to produce Paracoccidioides spp. antigens
    (2023) FERNANDES-BERALDO, Karolina Rosa; FREITAS-XAVIER, Roseli Santos de; PARDINI-VICENTINI, Adriana
    Introduction. 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 degrees C +/- 1 degrees C, on modified Fava-Netto agar, according to Freitas et al. (2018). Paracoccidioides lutzii antigens were obtained after 5, 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.
  • article
    Fast and cost-effective protocol for the production of Paracoccidioides spp. antigens
    (2023) FERNANDES-BERALDO, Karolina Rosa; FREITAS-XAVIER, Roseli Santos de; VICENTINI, Adriana Pardini
    Introduction: Existing methods for the production of Paracoccidioides spp. antigens are problematic considering standardization, specificity, stability, repeatability, and reproducibility. Objective: To optimize the methodology for producing Paracoccidioides spp. antigens and to evaluate its applicability in paracoccidioidomycosis immunodiagnosis. Materials and methods: The antigens were obtained from isolates of Paracoccidioides lutzii (01, 66 and 8334), Paracoccidioides brasiliensis sensu stricto (113) and Paracoccidioides restripiensis (B-339) grown at 36 degrees +/- 1 degrees C, on modified Fava-Netto agar, according Freitas et al. (2018). The P. lutzii antigens were obtained after 5, 10 and 20 days of culture and those of P. brasiliensis and P. restripiensis, after 10 days. The antigens were evaluated in natura, 10 and 20 times concentrated. The antigenic capacity was evaluated using the double immunodiffusion assay against serum samples from patients with paracoccidioidomycosis, histoplasmosis, aspergillosis and from random blood donors. Results: Cross-reactivity between Paracoccidioides spp. antigens was observed when P. brasiliensis and P. restrepiensis antigens were evaluated against polyclonal antibody anti-P. lutzii and when P. lutzii antigens were evaluated against polyclonal antibody anti-P. brasiliensis. There was no cross-reactivity against polyclonal antibodies anti-Histoplasma capsulatum and anti-Aspergillus fumigatus and random blood donors. The proposed protocol allowed the production of gender-specific, stable, repeatable and reproducible antigens, in a shorter cultivation time and at a lower cost. Conclusion: The proposed protocol allowed obtaining gender-specific antigens and can be developed and reproduced in all laboratories in Brazil and South America, where paracoccidioidomycosis is neglected, contributing to the early diagnosis, especially in endemic regions, regardless of the species.