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

Autor(es)

Roberto, Thiago Nunes ^[1] ; Rodrigues, Anderson Messias ^[1] ; Hahn, Rosane Christine ^[2] ; de Camargo, Zoilo Pires ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, Cellular Biol Div, Dept Microbiol Immunol & Parasitol, Sao Paulo - Brazil

^[2] Univ Fed Mato Grosso UFMT, Nucl Doencas Infecciosas & Trop, Cuiaba, MT - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Medical Mycology; v. 54, n. 3, p. 240-247, MAR 2016.
• Citações Web of Science: 10

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. (AU)

• Processo FAPESP: 11/07350-1 - Biologia do complexo Sporothrix schenckii: Análise de marcadores moleculares e proteômicos e o papel de moléculas antigênicas e antifúngicas na esporotricose
• Beneficiário: Anderson Messias Rodrigues
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 13/05405-9 - Análise da diversidade genética em agentes da paracoccidioidomicose usando marcadores AFLP (Amplified fragment length polymorphism)
• Beneficiário: Thiago Nunes Roberto
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 09/54024-2 - Biologia molecular e proteômica de fungos de interesse médico: Paracoccidioides brasiliensis e Sporothrix schenckii
• Beneficiário: Zoilo Pires de Camargo
• Modalidade de apoio: Auxílio à Pesquisa - Temático