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AutorLuiz, Raul Leal Faria
AutorMenezes, Rodrigo Caldas
AutorPereira, Sandro Antonio
AutorOliveira, Raquel de Vasconcellos Carvalhaes de
AutorOliveira, Manoel Marques Evangelista
Fecha de acceso2022-03-25T20:39:48Z
Fecha de disponibilización2022-03-25T20:39:48Z
Fecha de publicación2022
ReferenciaLUIZ, Raul Leal Faria et al. Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocols. Frontiers in veterinary science, v. 8, p. 1-10, 2022pt_BR
ISSN2297-1769pt_BR
URIhttps://www.arca.fiocruz.br/handle/icict/51885
Idiomaengpt_BR
EditorFrontiers Mediapt_BR
Derechos de autoropen accesspt_BR
TítuloNested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocolspt_BR
Tipo del documentoArticlept_BR
DOI10.3389/fvets.2021.755897
Resumen en InglésSporotrichosis is a chronic, cosmopolitan granulomatous mycosis that affects humans and animals. The infection is caused by the dimorphic fungi Sporothrix sp. The aims of the present study were to evaluate, standardize and validate a nested PCR technique using two DNA purification kits for the extraction of DNA from formalin fixed and paraffin-embedded tissues (FFPE) for Sporothrix sp. detection. FFPE mycological culture pellet samples of different Sporothrix species (S. chilensis, S. mexicana, S. pallida, S. globosa, S. brasiliensis and S. schenckii) were used as positive controls and clinical FFPE tissue samples of animals positive for Cryptococcus sp., Leishmania infantum and Histoplasma sp. were used as negative controls. Ten clinical FFPE skin samples from cats with sporotrichosis were used to validate the nested PCR. These samples were cut into two distinct paraffin sectioning protocols (5 and 16 μm thick). The paraffin sections were subjected to two different DNA extraction kits (chemical and thermal extractions). A nested PCR was performed on the extracted DNA to identify the genus Sporothrix. The chemical extraction protocol with the 5 μm thick paraffin section was more effective in extracting DNA from Sporothrix sp. from FFPE samples and the nested PCR technique showed the highest sensitivities (100% in the positive controls and of 50% in the skin samples of cats) and specificity (100%). Therefore, the nested PCR using this protocol has great potential to be applied in Sporothrix sp. diagnosis in FFPE samples of cats.pt_BR
AfiliaciónFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Taxonomia, Bioquímica e Bioprospecção de Fungos. Rio de Janeiro, RJ, Brasil.pt_BR
AfiliaciónFundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ, Brasil.pt_BR
AfiliaciónFundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ, Brasil.pt_BR
AfiliaciónFundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Epidemiologia Clínica. Rio de Janeiro, RJ, Brasil.pt_BR
AfiliaciónFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Taxonomia, Bioquímica e Bioprospecção de Fungos. Rio de Janeiro, RJ, Brasil.pt_BR
Palavras clave en InglêsDNA extractionpt_BR
Palavras clave en InglêsFFPE samplespt_BR
Palavras clave en InglêsSporothrix sp.pt_BR
Palavras clave en InglêsCatspt_BR
Palavras clave en InglêsMolecular diagnosispt_BR


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