Molecular epidemiology of Paracoccidioides brasiliensis. Preliminary report of the Argentinian ?Paraguayan Multicentric Study of Paracoccidioides and Paracoccidioidomycosis

SOSA, MARÍA DE LOS ÁNGELES; CATTANA MARÍA E.; ROJAS, FLORENCIA D.; FERNANDEZ MARIANA S.; GIUSIANO, GUSTAVO E.; NEGRONI, RICARDO; ARECHAVALA, ALICIA; SANTISO, GABRIELA; PARACOCCIDIOIDES AND PARACOCCIDOIDOMYCOSIS MULTICENTRIC STUDY

Foz de Iguazú

Workshop; XIV International Meeting on Paracoccidioidomycosis; 2017

Sociedade Brasileira de Microbiologia

Paracoccidioidomycosis (PCM) is the most important systemic fungal infection in Latin America. Primary pathogenic dimorphic fungi Paracoccidioides brasiliensis and P. lutzii are the etiological agents. Genetic diversity between phylogenetic species was reported. P. brasiliensis contains a complex of at least four different cryptic species (S1, PS2, PS3 and PS4). A great number of phenotypes may overlap in these related groups, discouraging the use of morphology alone for species recognition.Since 2014 an Argentinian-Paraguayan Multicentric study of Paracoccidioides and Paracoccidioidomicosis we have been conducting.In the present study, using PCR-RFLP of the alpha-tubulin (TUB1) gene, 35 strains were processed in order to identifying cryptic Paracoccidioides spp.DNA was extracted from 14-day colonies using boiling?thermic shock combination described by Giusiano et al.Alpha-tubulin gene fragments were amplified using primers α-Tub F (5-CTG GGA GGT ATG ATA ACACTG C-3) and α-Tub R (5-CGT CGG GCT ATT CAGATT TAA G-3) described by Roberto et al.A portion of the TUB1 gene was amplified and double digested in vitro using Bcl I and Msp I endonucleases, which generates four different electrophoretic patterns corresponding to the four main genetic groups: S1, PS2, and PS3 of P. brasiliensis and P. lutzii.As references, 11 strains previously sequenced and classified by WGS were used. P. brasiliensis S1 (n=33; 94.3 %) was the major group recognized followed by PS2 (n = 2; 5.7 %).Except for one strain (PbP99) misidentified by TUB1-RFLP, a good agreement with WGS was observed.TUB1-RFLP is a fast and inexpensive tool for identifying Paracoccidioides spp., which can be used as a screening method to apply in molecular epidemiological studies of paracoccidioidomycosis.