Identifying Paracoccidioides phylogenetic species by PCR-RFLP of the alpha-tubulin gene

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dc.citation.issue3
dc.citation.volume54
dc.contributor.authorRoberto, Thiago Nunes [UNIFESP]
dc.contributor.authorRodrigues, Anderson Messias [UNIFESP]
dc.contributor.authorHahn, Rosane Christine
dc.contributor.authorCamargo, Zoilo Pires de [UNIFESP]
dc.coverageOxford
dc.date.accessioned2020-08-21T17:00:09Z
dc.date.available2020-08-21T17:00:09Z
dc.date.issued2016
dc.description.abstractParacoccidioidomycosis 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.en
dc.description.affiliationUniv Fed Sao Paulo, Cellular Biol Div, Dept Microbiol Immunol & Parasitol, Sao Paulo, Brazil
dc.description.affiliationUniv Fed Mato Grosso UFMT, Nucl Doencas Infecciosas & Trop, Cuiaba, MT, Brazil
dc.description.affiliationUnifespUniv Fed Sao Paulo, Cellular Biol Div, Dept Microbiol Immunol & Parasitol, Sao Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.description.sponsorshipIDFAPESP: 2013/05405-9
dc.description.sponsorshipIDFAPESP: 2011/07350-1
dc.description.sponsorshipIDFAPESP: 2009/54024-2
dc.format.extent240-247
dc.identifierhttp://dx.doi.org/10.1093/mmy/myv083
dc.identifier.citationMedical Mycology. Oxford, v. 54, n. 3, p. 240-247, 2016.
dc.identifier.doi10.1093/mmy/myv083
dc.identifier.issn1369-3786
dc.identifier.urihttps://repositorio.unifesp.br/handle/11600/57888
dc.identifier.wosWOS:000372105200003
dc.language.isoeng
dc.publisherOxford Univ Press
dc.relation.ispartofMedical Mycology
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectParacoccidioidesen
dc.subjectParacoccidioidomycosisen
dc.subjectPCR-RFLPen
dc.subjectMolecular identificationen
dc.subjectAlpha-tubulinen
dc.titleIdentifying Paracoccidioides phylogenetic species by PCR-RFLP of the alpha-tubulin geneen
dc.typeinfo:eu-repo/semantics/article
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