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      Diagnóstico micológico de paracoccidioidomicosis en un hospital de área no endémica: metodología clásica y molecular Translated title: Mycological diagnosis of paracoccidioidomycosis in a hospital from a nonendemic area: classical and molecular methods

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          Resumen

          Introducción.

          La paracoccidioidomicosis es una micosis sistémica y endémica en Latinoamérica. El cambio climático y el movimiento migratorio del huésped enfatizan la necesidad de optimizar el diagnóstico de esta infección.

          Objetivo.

          Evaluar la implementación de la detección de ADN de Paracoccidioides spp. al diagnóstico micológico de pacientes con sospecha de paracoccidioidomicosis.

          Materiales y métodos.

          Estudio retrospectivo con datos de laboratorio de pacientes con sospecha de paracoccidioidomicosis en un hospital de área no endémica.

          Resultados.

          Se analizaron los resultados de las muestras de 19 pacientes con sospecha clínica de paracoccidioidomicosis. El 90 % de los pacientes había nacido o visitado un área endémica de esta micosis en Latinoamérica. En 14 pacientes varones adultos se confirmó paracoccidioidomicosis por diagnóstico convencional. El examen directo fue positivo en 12 pacientes con enfermedad comprobada y en 4 de ellos se obtuvo crecimiento del hongo. Se detectaron anticuerpos contra Paracoccidioides spp. en ocho pacientes con la enfermedad. Se realizó PCR anidada con muestras de 14 pacientes para detectar ADN de Paracoccidioides spp. En 9 de los 10 pacientes con diagnóstico convencional de paracoccidioidomicosis se obtuvo una prueba de PCR positiva.

          Conclusiones.

          La implementación de técnicas moleculares para detectar ADN de Paracoccidioides spp. complementa el diagnóstico convencional de paracoccidioidomicosis y permite instaurar el tratamiento antifúngico, sobre todo en los casos clínicos donde no se observa la presencia del hongo en las muestras clínicas. La migración actual de poblaciones humanas dificulta el diagnóstico de paracoccidioidiomicosis y otras infecciones endémicas, por lo que se requiere optimizar el diagnostico micológico en los laboratorios clínicos para tratar pacientes con este tipo micosis desatendida.

          Abstract

          Introduction.

          Paracoccidioidomycosis is a systemic mycosis endemic in Latin America. Climate change and host migration emphasize the need to optimize this infection diagnosis.

          Objective.

          To evaluate the implementation of Paracoccidioides spp. DNA detection in the mycological diagnosis of patients with suspected paracoccidioidomycosis.

          Materials and methods.

          It is a retrospective study with laboratory data from patients with clinical suspicion of paracoccidioidomycosis, who consulted a university hospital from a non-endemic area.

          Results.

          We analyzed the laboratory results of samples from 19 patients with suspected paracoccidioidomycosis. Seventeen out of 19 patients were born in or had visited an endemic area in Latin America. Fourteen adult male patients were confirmed to have paracoccidioidomycosis by conventional diagnosis: the direct examination was positive in 12 samples while fungal growth was found only in 4. Anti- Paracoccidioides spp. antibodies were detected in 10 patients, 8 of them with proven paracoccidioidomycosis. Nested PCR for Paracoccidioides spp. detection was performed on clinical samples from 14 patients, and positive results were obtained for 9 out of 10 patients with the conventional diagnosis of paracoccidioidomycosis.

          Conclusions.

          The incorporation of molecular techniques to detect Paracoccidioides spp. DNA complements the conventional diagnosis of paracoccidioidomycosis. This tool allows the prescription of antifungal treatment in those cases where the fungus is not observed in the clinical samples. Current human migrations difficult the mycological diagnosis of paracoccidioidomycosis and other fungal infections. For this reason, it is necessary to improve mycological diagnosis in clinical laboratories to adequately treat patients with this neglected mycosis.

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          Most cited references44

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          Brazilian guidelines for the clinical management of paracoccidioidomycosis

          Abstract Paracoccidioidomycosis is a systemic fungal disease occurring in Latin America that is associated with rural environments and agricultural activities. However, the incidence and prevalence of paracoccidiodomycosis is underestimated because of the lack of compulsory notification. If paracoccidiodomycosis is not diagnosed and treated early and adequately, the endemic fungal infection could result in serious sequelae. While the Paracoccidioides brasiliensis ( P. brasiliensis ) complex has been known to be the causal agent of paracoccidiodomycosis, a new species, Paracoccidioides lutzii ( P. lutzii ), has been reported in Rondônia, where the disease has reached epidemic levels, and in the Central West and Pará. Accurate diagnoses and availability of antigens that are reactive with the patients’ sera remain significant challenges. Therefore, the present guidelines aims to update the first Brazilian consensus on paracoccidioidomycosis by providing evidence-based recommendations for bedside patient management. This consensus summarizes etiological, ecoepidemiological, molecular epidemiological, and immunopathological data, with emphasis on clinical, microbiological, and serological diagnosis and management of clinical forms and sequelae, as well as in patients with comorbidities and immunosuppression. The consensus also includes discussion of outpatient treatments, severe disease forms, disease prevalence among special populations and resource-poor settings, a brief review of prevention and control measures, current challenges and recommendations.
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            Epidemiology of endemic systemic fungal infections in Latin America.

            Although endemic mycoses are a frequent health problem in Latin American countries, clinical and epidemiological data remain scarce and fragmentary. These mycoses have a significant impact on public health, and early diagnosis and appropriate treatment remain important. The target population for endemic disease in Latin America is mostly represented by low-income rural workers with limited access to a public or private health system. Unfortunately, diagnostic tools are not widely available in medical centers in Latin America; consequently, by the time patients are diagnosed with fungal infection, many are already severely ill. Among immunocompromised patients, endemic mycoses usually behave as opportunistic infections causing disseminated rather than localized disease. This paper reviews the epidemiology of the most clinically significant endemic mycoses in Latin America: paracoccidioidomycosis, histoplasmosis, and coccidioidomycosis. The burdens of disease, typically affected populations, and clinical outcomes also are discussed.
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              Species boundaries in the human pathogen Paracoccidioides.

              The use of molecular taxonomy for identifying recently diverged species has transformed the study of speciation in fungi. The pathogenic fungus Paracoccidioides spp has been hypothesized to be composed of five phylogenetic species, four of which compose the brasiliensis species complex. Nuclear gene genealogies support this divergence scenario, but mitochondrial loci do not; while all species from the brasiliensis complex are differentiated at nuclear coding loci, they are not at mitochondrial loci. We addressed the source of this incongruity using 11 previously published gene fragments, 10 newly-sequenced nuclear non-coding loci, and 10 microsatellites. We hypothesized and further demonstrated that the mito-nuclear incongruence in the brasiliensis species complex results from interspecific hybridization and mitochondrial introgression, a common phenomenon in eukaryotes. Additional population genetic analyses revealed possible nuclear introgression but much less than that seen in the mitochondrion. Our results are consistent with a divergence scenario of secondary contact and subsequent mitochondrial introgression despite the continued persistence of species boundaries. We also suggest that yeast morphology slightly-but significantly-differs across all five Paracoccidioides species and propose to elevate four of these phylogenetic species to formally described taxonomic species.
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                Author and article information

                Journal
                Biomedica
                Biomedica
                bio
                Biomédica
                Instituto Nacional de Salud
                0120-4157
                2590-7379
                31 August 2023
                August 2023
                : 43
                : Suppl 1
                : 132-143
                Affiliations
                [1 ] originalLaboratorio de Micología, Hospital de Clínicas “José de San Martín”, Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires, Argentina normalizedUniversidad de Buenos Aires orgnameUniversidad de Buenos Aires Ciudad Autónoma de Buenos Aires, Argentina
                [2 ] originalServicio Micosis Profundas, Departamento de Micología, INEI-ANLIS “Dr. Carlos G. Malbrán”, Ciudad Autónoma de Buenos Aires, Argentina orgnameDepartamento de Micología, INEI-ANLIS “Dr. Carlos G. Malbrán” Ciudad Autónoma de Buenos Aires, Argentina
                [3 ] originalDepartamento de Micología, INEI-ANLIS “Dr. Carlos G. Malbrán”, Ciudad Autónoma de Buenos Aires, Argentina orgnameDepartamento de Micología, INEI-ANLIS “Dr. Carlos G. Malbrán” Ciudad Autónoma de Buenos Aires, Argentina
                Author notes
                [* ] Correspondencia: Norma B. Fernández, Laboratorio de Micología, Hospital de Clínicas “José de San Martín”, Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires, Argentina Teléfono: (54 11) 3448 0263 normafer38@ 123456gmail.com

                Todos los autores participaron en todas las etapas del estudio.

                Conflicto de intereses: Ninguno para declarar.

                Article
                10.7705/biomedica.6888
                10569774
                37721907
                8a15671f-37ff-4329-b57c-946887669853

                Este es un artículo publicado en acceso abierto bajo una licencia Creative Commons

                History
                : 01 February 2023
                : 02 May 2023
                : 05 May 2023
                Page count
                Figures: 0, Tables: 1, Equations: 0, References: 44, Pages: 12
                Categories
                Artículo Original

                paracoccidioides,paracoccidioidomicosis,técnicas de diagnóstico molecular,adn,micosis,paracoccidioidomycosis,molecular diagnostic techniques,dna,mycoses

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