| Author | Luiz, Raul Leal Faria | |
| Author | Menezes, Rodrigo Caldas | |
| Author | Pereira, Sandro Antonio | |
| Author | Oliveira, Raquel de Vasconcellos Carvalhaes de | |
| Author | Oliveira, Manoel Marques Evangelista | |
| Access date | 2022-09-27T13:50:28Z | |
| Available date | 2022-09-27T13:50:28Z | |
| Document date | 2022 | |
| Citation | LUIZ, Raul Leal Faria et al. Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DN11A Extraction Protocols. Frontiers in Veterinary Science, V.8, Article 755897, 10p. January 2022. | en_US |
| ISSN | 2297-1769 | en_US |
| URI | https://www.arca.fiocruz.br/handle/icict/54879 | |
| Language | eng | en_US |
| Publisher | Frontiers Midia | en_US |
| Rights | open access | en_US |
| Title | Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocols | en_US |
| Type | Article | en_US |
| DOI | 10.3389/fvets.2021.755897 | |
| Abstract | 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. | en_US |
| Affilliation | Instituto Oswaldo Cruz. Laboratório. Laboratório de Taxonomia, Bioquímica e Bioprospecção de Fungos. RJ, Brasil. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ, Brasil. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em Dermatozoonoses em Animais Domésticos. Rio de Janeiro, RJ, Brasil. | en_US |
| Affilliation | Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Epidemiologia Clínica. Rio de Janeiro, RJ, Brasil. | en_US |
| Affilliation | Instituto Oswaldo Cruz. Laboratório. Laboratório de Taxonomia, Bioquímica e Bioprospecção de Fungos. RJ, Brasil. | en_US |
| Subject | Sporothrix sp | en_US |
| Subject | FFPE samples | en_US |
| Subject | DNA extraction | en_US |
| Subject | Molecular diagnosis | en_US |
| Subject | Cats | en_US |
