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https://www.arca.fiocruz.br/handle/icict/38795
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ArticleCopyright
Open access
Embargo date
2020-12-23
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- INI - Artigos de Periódicos [3397]
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UTILITY OF REAL-TIME PCR FOR THE DETECTION OF PARACOCCIDIOIDES BRASILIENSIS DNA IN THE DIAGNOSIS OF IMPORTED PARACOCCIDIOIDOMYCOSIS
Author
Affilliation
Instituto de Salud Carlos III. Servicio de Micología. Majadahonda, Spain.
Hospital Clinico San Carlos. Madrid, Spain.
Hospital Carlos III. Madrid, Spain.
Instituto de Salud Carlos III. Servicio de Micología. Majadahonda, Spain.
Hospital Clinico San Carlos. Madrid, Spain.
Fundação Oswaldo Cruz. Instituto de Pesquisa Clinica Evandro Chagas. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto de Pesquisa Clinica Evandro Chagas. Rio de Janeiro, RJ, Brasil.
Instituto de Salud Carlos III. Servicio de Micología. Majadahonda, Spain.
Instituto de Salud Carlos III. Servicio de Micología. Majadahonda, Spain.
Hospital Clinico San Carlos. Madrid, Spain.
Hospital Carlos III. Madrid, Spain.
Instituto de Salud Carlos III. Servicio de Micología. Majadahonda, Spain.
Hospital Clinico San Carlos. Madrid, Spain.
Fundação Oswaldo Cruz. Instituto de Pesquisa Clinica Evandro Chagas. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto de Pesquisa Clinica Evandro Chagas. Rio de Janeiro, RJ, Brasil.
Instituto de Salud Carlos III. Servicio de Micología. Majadahonda, Spain.
Instituto de Salud Carlos III. Servicio de Micología. Majadahonda, Spain.
Abstract
An increase in immigration from endemic regions has resulted in a number of cases of paracoccidioidomycosis (PCM) being imported into Spain. A molecular diagnostic technique based on real-time PCR was developed for the detection of Paracoccidioides brasiliensis DNA in both culture and patients’ clinical samples. A Molecular Beacon probe was used, labelled with FAM and directed at the ITS1 region of ribosomic DNA. The detection limit of the technique developed was 1 fg of fungal DNA per μl of sample. This procedure proved to be very reproducible and specifi c. The technique was tested with cultures of 12 clinical strains and on samples from two patients with proven PCM. Real-time PCR was positive for all the culture strains, as well as those from both patients. By samples, the technique was positive in sputum and tissue biopsies but less useful on blood samples. Samples were analyzed several months after patient treatment, detecting a small amount of fungal DNA in one respiratory sample. This technique of real-time PCR is a sensitive method for rapid diagnosis of paracoccidioidomycosis and could serve to monitor patients after treatment has begun.
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