Identifying Paracoccidioides phylogenetic species by PCR-RFLP of the alpha-tubulin gene
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | https://repositorio.unifesp.br/handle/11600/57888 http://dx.doi.org/10.1093/mmy/myv083 |
Resumo: | Paracoccidioidomycosis is an important systemic fungal infection that occurs throughout Latin America. The etiological agents comprise a species complex that includes two major groups: P. brasiliensis (including subgroups S1, PS2, and PS3) and P. lutzii. A great number of phenotypes may overlap, especially among closely related groups, discouraging the use ofmorphology alone for species recognition. To overcome this problem, here we propose identifying cryptic Paracoccidioides spp. using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the alpha-tubulin (TUB1) gene. In silico analysis of 90 TUB1 sequences led to the identification of two restriction enzymes with the potential to identify Paracoccidioides: Bcl I and MspI. A portion of the TUB1 gene was amplified and double digested in vitro with the Bcl I and MspI endonucleases, which generated four different electrophoretic patterns corresponding to the four main genetic groups: S1, PS2, and PS3 of P. brasiliensis and P. lutzii. The major P. brasiliensis group recognized was S1 (n = 17; 42.5%), followed by PS2 (n = 9; 22.5%) and PS3 (n = 6; 15%). A total of eight (20%) P. lutzii isolates were identified, mainly from mid western Brazil. Our data revealed that TUB1-RFLP is an efficient, fast, and inexpensive tool for identifying Paracoccidioides spp., which may be directly applied to the molecular epidemiological studies of paracoccidioidomycosis. |
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Roberto, Thiago Nunes [UNIFESP]Rodrigues, Anderson Messias [UNIFESP]Hahn, Rosane ChristineCamargo, Zoilo Pires de [UNIFESP]2020-08-21T17:00:09Z2020-08-21T17:00:09Z2016Medical Mycology. Oxford, v. 54, n. 3, p. 240-247, 2016.1369-3786https://repositorio.unifesp.br/handle/11600/57888http://dx.doi.org/10.1093/mmy/myv08310.1093/mmy/myv083WOS:000372105200003Paracoccidioidomycosis is an important systemic fungal infection that occurs throughout Latin America. The etiological agents comprise a species complex that includes two major groups: P. brasiliensis (including subgroups S1, PS2, and PS3) and P. lutzii. A great number of phenotypes may overlap, especially among closely related groups, discouraging the use ofmorphology alone for species recognition. To overcome this problem, here we propose identifying cryptic Paracoccidioides spp. using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the alpha-tubulin (TUB1) gene. In silico analysis of 90 TUB1 sequences led to the identification of two restriction enzymes with the potential to identify Paracoccidioides: Bcl I and MspI. A portion of the TUB1 gene was amplified and double digested in vitro with the Bcl I and MspI endonucleases, which generated four different electrophoretic patterns corresponding to the four main genetic groups: S1, PS2, and PS3 of P. brasiliensis and P. lutzii. The major P. brasiliensis group recognized was S1 (n = 17; 42.5%), followed by PS2 (n = 9; 22.5%) and PS3 (n = 6; 15%). A total of eight (20%) P. lutzii isolates were identified, mainly from mid western Brazil. Our data revealed that TUB1-RFLP is an efficient, fast, and inexpensive tool for identifying Paracoccidioides spp., which may be directly applied to the molecular epidemiological studies of paracoccidioidomycosis.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Univ Fed Sao Paulo, Cellular Biol Div, Dept Microbiol Immunol & Parasitol, Sao Paulo, BrazilUniv Fed Mato Grosso UFMT, Nucl Doencas Infecciosas & Trop, Cuiaba, MT, BrazilUniv Fed Sao Paulo, Cellular Biol Div, Dept Microbiol Immunol & Parasitol, Sao Paulo, BrazilFAPESP: 2013/05405-9FAPESP: 2011/07350-1FAPESP: 2009/54024-2Web of Science240-247engOxford Univ PressMedical MycologyParacoccidioidesParacoccidioidomycosisPCR-RFLPMolecular identificationAlpha-tubulinIdentifying Paracoccidioides phylogenetic species by PCR-RFLP of the alpha-tubulin geneinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleOxford543info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP11600/578882021-08-11 11:40:52.098metadata only accessoai:repositorio.unifesp.br:11600/57888Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652021-08-11T14:40:52Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Identifying Paracoccidioides phylogenetic species by PCR-RFLP of the alpha-tubulin gene |
title |
Identifying Paracoccidioides phylogenetic species by PCR-RFLP of the alpha-tubulin gene |
spellingShingle |
Identifying Paracoccidioides phylogenetic species by PCR-RFLP of the alpha-tubulin gene Roberto, Thiago Nunes [UNIFESP] Paracoccidioides Paracoccidioidomycosis PCR-RFLP Molecular identification Alpha-tubulin |
title_short |
Identifying Paracoccidioides phylogenetic species by PCR-RFLP of the alpha-tubulin gene |
title_full |
Identifying Paracoccidioides phylogenetic species by PCR-RFLP of the alpha-tubulin gene |
title_fullStr |
Identifying Paracoccidioides phylogenetic species by PCR-RFLP of the alpha-tubulin gene |
title_full_unstemmed |
Identifying Paracoccidioides phylogenetic species by PCR-RFLP of the alpha-tubulin gene |
title_sort |
Identifying Paracoccidioides phylogenetic species by PCR-RFLP of the alpha-tubulin gene |
author |
Roberto, Thiago Nunes [UNIFESP] |
author_facet |
Roberto, Thiago Nunes [UNIFESP] Rodrigues, Anderson Messias [UNIFESP] Hahn, Rosane Christine Camargo, Zoilo Pires de [UNIFESP] |
author_role |
author |
author2 |
Rodrigues, Anderson Messias [UNIFESP] Hahn, Rosane Christine Camargo, Zoilo Pires de [UNIFESP] |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Roberto, Thiago Nunes [UNIFESP] Rodrigues, Anderson Messias [UNIFESP] Hahn, Rosane Christine Camargo, Zoilo Pires de [UNIFESP] |
dc.subject.eng.fl_str_mv |
Paracoccidioides Paracoccidioidomycosis PCR-RFLP Molecular identification Alpha-tubulin |
topic |
Paracoccidioides Paracoccidioidomycosis PCR-RFLP Molecular identification Alpha-tubulin |
description |
Paracoccidioidomycosis is an important systemic fungal infection that occurs throughout Latin America. The etiological agents comprise a species complex that includes two major groups: P. brasiliensis (including subgroups S1, PS2, and PS3) and P. lutzii. A great number of phenotypes may overlap, especially among closely related groups, discouraging the use ofmorphology alone for species recognition. To overcome this problem, here we propose identifying cryptic Paracoccidioides spp. using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the alpha-tubulin (TUB1) gene. In silico analysis of 90 TUB1 sequences led to the identification of two restriction enzymes with the potential to identify Paracoccidioides: Bcl I and MspI. A portion of the TUB1 gene was amplified and double digested in vitro with the Bcl I and MspI endonucleases, which generated four different electrophoretic patterns corresponding to the four main genetic groups: S1, PS2, and PS3 of P. brasiliensis and P. lutzii. The major P. brasiliensis group recognized was S1 (n = 17; 42.5%), followed by PS2 (n = 9; 22.5%) and PS3 (n = 6; 15%). A total of eight (20%) P. lutzii isolates were identified, mainly from mid western Brazil. Our data revealed that TUB1-RFLP is an efficient, fast, and inexpensive tool for identifying Paracoccidioides spp., which may be directly applied to the molecular epidemiological studies of paracoccidioidomycosis. |
publishDate |
2016 |
dc.date.issued.fl_str_mv |
2016 |
dc.date.accessioned.fl_str_mv |
2020-08-21T17:00:09Z |
dc.date.available.fl_str_mv |
2020-08-21T17:00:09Z |
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 |
Medical Mycology. Oxford, v. 54, n. 3, p. 240-247, 2016. |
dc.identifier.uri.fl_str_mv |
https://repositorio.unifesp.br/handle/11600/57888 http://dx.doi.org/10.1093/mmy/myv083 |
dc.identifier.issn.none.fl_str_mv |
1369-3786 |
dc.identifier.doi.none.fl_str_mv |
10.1093/mmy/myv083 |
dc.identifier.wos.none.fl_str_mv |
WOS:000372105200003 |
identifier_str_mv |
Medical Mycology. Oxford, v. 54, n. 3, p. 240-247, 2016. 1369-3786 10.1093/mmy/myv083 WOS:000372105200003 |
url |
https://repositorio.unifesp.br/handle/11600/57888 http://dx.doi.org/10.1093/mmy/myv083 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Medical Mycology |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
240-247 |
dc.coverage.none.fl_str_mv |
Oxford |
dc.publisher.none.fl_str_mv |
Oxford Univ Press |
publisher.none.fl_str_mv |
Oxford Univ Press |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UNIFESP instname:Universidade Federal de São Paulo (UNIFESP) instacron:UNIFESP |
instname_str |
Universidade Federal de São Paulo (UNIFESP) |
instacron_str |
UNIFESP |
institution |
UNIFESP |
reponame_str |
Repositório Institucional da UNIFESP |
collection |
Repositório Institucional da UNIFESP |
repository.name.fl_str_mv |
Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP) |
repository.mail.fl_str_mv |
|
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1802764109799751680 |