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Genotyping of Paracoccidioides brasiliensis from different samples of paracoccidioidomycosis patients treated at Botucatu Medical School -UNESP

Grant number: 13/24877-9
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): March 01, 2014
Effective date (End): February 28, 2018
Field of knowledge:Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
Principal researcher:Luciane Alarcão Dias-Melicio
Grantee:Luciana Bonome Zeminian de Oliveira
Home Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Paracoccidioidomycosis is a systemic fungal infection and its etiologic agent is Paracoccidioides brasiliensis (P. brasiliensis). Using concordance of gene genealogies was detected three Cryptic species: S2, PS2 and PS3. From multilocus phylogenetic analysis were demonstrated the existence of a group genetically divergent from the isolates of P. brasiliensis from the complex S1, PS2 and PS3, the Pb01-like, originating of de Midwest region of Brazil which constitute a fourth cryptic specie, Paracoccidioides lutzii. Genotyping the samples of the line Paracoccidoides may contribute to clinical and epidemiological studies of paracoccidioidomycosis. Thus, with all this knowledge, it's very interesting an assessment of the different species in samples collected from patients treated in Botucatu. This study has as its goal the evaluation of the speciation of the Paracoccidioides in samples embedded in paraffin of patients treated at Botucatu Medical School, UNESP. For this, it's proposed a DNA extraction of the embedded samples, followed by a PCR and the sequencing of the ITS region of the Paracoccidioides genetic code to distinguish P. brasiliensis from P. lutzii. Then, for the speciation of the isolates S1, PS2 e PS3 it's proposed a PCR Real Time technique called Taqman for the gp43 and ARF regions followed by a sequencing of this same regions, for confirmation of the data. The sequences will be aligned and analyzed with the software MEGA 5.2 (Kumar et al, 2012) using the algorithm of alignment MUSCLE (Edgar, 2004). The sequences will be compared to the sequences already deposited in GenBank. The best evolutive model will be calculated with the new sequences and the phylogenetic tree will be drawn by the methods of Neighbor Joining and maximum-likelihood estimation, using as statistic support method 1000 copies of bootstrap.

Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
OLIVEIRA, Luciana Bonome Zeminian de. Genotipagem do Paracoccidioides brasiliensis de diferentes amostras de pacientes atendidos no Hospital das Clínicas da Faculdade de Medicina de Botucatu. 2018. Doctoral Thesis - Universidade Estadual Paulista "Júlio de Mesquita Filho" Faculdade de Medicina..

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