Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocols

Detalhes bibliográficos
Autor(a) principal: Luiz, Raul Leal Faria
Data de Publicação: 2022
Outros Autores: Menezes, Rodrigo Caldas, Pereira, Sandro Antonio, Oliveira, Raquel de Vasconcellos Carvalhaes de, Oliveira, Manoel Marques Evangelista
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/51885
Resumo: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Taxonomia, Bioquímica e Bioprospecção de Fungos. Rio de Janeiro, RJ, Brasil.
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spelling Luiz, Raul Leal FariaMenezes, Rodrigo CaldasPereira, Sandro AntonioOliveira, Raquel de Vasconcellos Carvalhaes deOliveira, Manoel Marques Evangelista2022-03-25T20:39:48Z2022-03-25T20:39:48Z2022LUIZ, 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, 20222297-1769https://www.arca.fiocruz.br/handle/icict/5188510.3389/fvets.2021.755897engFrontiers MediaNested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocolsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Taxonomia, Bioquímica e Bioprospecção de Fungos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Epidemiologia Clínica. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Taxonomia, Bioquímica e Bioprospecção de Fungos. Rio de Janeiro, RJ, Brasil.Sporotrichosis 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.DNA extractionFFPE samplesSporothrix sp.CatsMolecular diagnosisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83099https://www.arca.fiocruz.br/bitstream/icict/51885/1/license.txt586c046dcfeef936e32f0323bb9a47c0MD51ORIGINALNested_Rodrigo_Menezes_etal_INI_2022.pdfNested_Rodrigo_Menezes_etal_INI_2022.pdfapplication/pdf744729https://www.arca.fiocruz.br/bitstream/icict/51885/2/Nested_Rodrigo_Menezes_etal_INI_2022.pdf391c239f4339cf761683d8ce5ec3c083MD52icict/518852022-03-25 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dc.title.pt_BR.fl_str_mv Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocols
title Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocols
spellingShingle Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocols
Luiz, Raul Leal Faria
DNA extraction
FFPE samples
Sporothrix sp.
Cats
Molecular diagnosis
title_short Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocols
title_full Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocols
title_fullStr Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocols
title_full_unstemmed Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocols
title_sort Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocols
author Luiz, Raul Leal Faria
author_facet Luiz, Raul Leal Faria
Menezes, Rodrigo Caldas
Pereira, Sandro Antonio
Oliveira, Raquel de Vasconcellos Carvalhaes de
Oliveira, Manoel Marques Evangelista
author_role author
author2 Menezes, Rodrigo Caldas
Pereira, Sandro Antonio
Oliveira, Raquel de Vasconcellos Carvalhaes de
Oliveira, Manoel Marques Evangelista
author2_role author
author
author
author
dc.contributor.author.fl_str_mv Luiz, Raul Leal Faria
Menezes, Rodrigo Caldas
Pereira, Sandro Antonio
Oliveira, Raquel de Vasconcellos Carvalhaes de
Oliveira, Manoel Marques Evangelista
dc.subject.en.pt_BR.fl_str_mv DNA extraction
FFPE samples
Sporothrix sp.
Cats
Molecular diagnosis
topic DNA extraction
FFPE samples
Sporothrix sp.
Cats
Molecular diagnosis
description Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Taxonomia, Bioquímica e Bioprospecção de Fungos. Rio de Janeiro, RJ, Brasil.
publishDate 2022
dc.date.accessioned.fl_str_mv 2022-03-25T20:39:48Z
dc.date.available.fl_str_mv 2022-03-25T20:39:48Z
dc.date.issued.fl_str_mv 2022
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv LUIZ, 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, 2022
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/51885
dc.identifier.issn.pt_BR.fl_str_mv 2297-1769
dc.identifier.doi.none.fl_str_mv 10.3389/fvets.2021.755897
identifier_str_mv LUIZ, 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, 2022
2297-1769
10.3389/fvets.2021.755897
url https://www.arca.fiocruz.br/handle/icict/51885
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Frontiers Media
publisher.none.fl_str_mv Frontiers Media
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/51885/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/51885/2/Nested_Rodrigo_Menezes_etal_INI_2022.pdf
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repository.name.fl_str_mv Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)
repository.mail.fl_str_mv repositorio.arca@fiocruz.br
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