Advanced search
Start date
Betweenand
Related content
(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

Full text
Author(s):
Roberto, Thiago Nunes [1] ; Rodrigues, Anderson Messias [1] ; Hahn, Rosane Christine [2] ; de Camargo, Zoilo Pires [1]
Total Authors: 4
Affiliation:
[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
Total Affiliations: 2
Document type: Journal article
Source: Medical Mycology; v. 54, n. 3, p. 240-247, MAR 2016.
Web of Science Citations: 10
Abstract

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)

FAPESP's process: 11/07350-1 - Biology of Sporothrix schenckii complex: Analysis of molecular and proteomics markers, and the role of antigenic and antifungal molecules in sporotrichosis
Grantee:Anderson Messias Rodrigues
Support Opportunities: Scholarships in Brazil - Doctorate
FAPESP's process: 13/05405-9 - Analysis of genetic diversity in agents of paracoccidioidomycosis using AFLP (Amplified fragment length polymorphism)
Grantee:Thiago Nunes Roberto
Support Opportunities: Scholarships in Brazil - Master
FAPESP's process: 09/54024-2 - Molecular biology and proteomics of medically interesting fungi: Paracoccidioides brasiliensis and Sporothrix schenckii
Grantee:Zoilo Pires de Camargo
Support Opportunities: Research Projects - Thematic Grants