XI International Meeting on Paracoccidioidomycosis

XI International Meeting on Paracoccidioidomycosis

Resume:106-2

Poster (Painel)

106-2	IDENTIFICATION OF Pb01-like (P. lutzii) AMONG Paracoccidioides spp. ISOLATES ON THE PREVIOUSLY CHARACTERIZED hsp70 GENE INDEL
Authors:	Fernanda Lourenã§o Alves (UFMG - Universidade Federal de Minas Gerais) ; Marjorie Mendes Marini (UNIFESP - Universidade Federal de SÃ£o Paulo) ; Patrãcia Silva Cisalpino (UFMG - Universidade Federal de Minas Gerais) ; Marco Aurã©lio Soares (UFMG - Universidade Federal de Minas Gerais)

Abstract

Paracoccidioides brasiliensis is the etiological agent of paracoccidioidomycosis, a systemic disease endemic to most Latin America, wich in Brazil represents the eighth leading cause of death among infectious and parasitic diseases.Recent studies suggest that the genus Paracoccidioides consists of at least four different phylogenetic species:P. brasiliensis S1, PS2 and PS3 and a fourth distinct lineage comprising fungal isolates referred as Pb01-like, now proposed to be a completely different new species, P. lutzii.This study aims to identify P. lutzii in different isolates of Paracoccidioides spp. Thirty-five isolates of Paracoccidioides spp. were used for DNA extraction and for subsequent Polymerase Chain Reaction (PCR) using primers previously described (HSPMMT1 and PLMMT1) that amplify a region of 400 bp from 5'-UTR to intron 1 of the hsp70 gene. PCR were carried out on 10 ng of DNA in a 20 μL reaction mixture containing: 10 mM Tris-HCl pH 9; 1.5 mM MgCl2, 50 mM KCl; 0.2 mM of each dNTP; 0.2 of each oligonucleotide and 1 unit of Taq DNA Polymerase. The cycling conditions were as follows: denaturing at 94•C for 5 min, anneling at 60•C for 1 min and extension at 72•C for 1 min. At the end of the 30th cycle, the heat-denaturing step was omitted and extension was allowed to proceed at 72•C for 5 min. The amplified product was visualized on 0.8% agarose gel stained with ethidium bromide.਀䄀㐀　　戀瀀戀愀渀搀眀愀猀瘀椀猀甀愀氀椀稀攀搀椀渀琀栀爀攀攀欀渀漀眀渀倀戀　㄀ⴀ㰀椀㸀氀椀欀攀㰀⼀椀㸀椀猀漀氀愀琀攀猀 ⠀倀戀　㄀Ⰰ ㄀㔀㜀㠀Ⰰ 䔀䐀　㄀⤀ 愀渀搀椀渀椀猀漀氀愀琀攀㘀㌀㈀㘀㔀Ⰰ 挀漀渀猀椀搀攀爀攀搀琀漀戀攀氀漀渀最琀漀瀀栀礀氀漀最攀渀攀琀椀挀猀瀀攀挀椀攀猀匀㄀⸀ 䤀渀琀栀攀洀愀樀漀爀椀琀礀漀昀椀猀漀氀愀琀攀猀眀攀爀攀漀戀猀攀爀瘀攀搀渀漀渀猀瀀攀挀椀昀椀挀戀愀渀搀猀漀昀㔀　　愀渀搀㘀　　戀瀀⸀吀栀攀瀀愀椀爀漀昀瀀爀椀洀攀爀猀眀愀猀甀猀攀昀甀氀椀渀椀搀攀渀琀椀昀礀椀渀最椀猀漀氀愀琀攀猀漀昀琀栀攀猀瀀攀挀椀攀猀㰀椀㸀倀⸀ 氀甀琀稀椀椀㰀⼀椀㸀Ⰰ 眀栀椀挀栀猀栀愀爀攀愀渀椀渀搀攀氀椀渀琀栀攀昀椀爀猀琀椀渀琀爀漀渀漀昀㰀椀㸀栀猀瀀㜀　㰀⼀椀㸀最攀渀攀⸀吀栀攀椀猀漀氀愀琀攀猀琀栀愀琀猀栀漀眀攀搀渀漀渀猀瀀攀挀椀昀椀挀戀愀渀搀猀愀渀搀琀栀攀椀猀漀氀愀琀攀㘀㌀㈀㘀㔀猀栀漀甀氀搀戀攀猀甀戀洀椀琀琀攀搀琀漀猀攀焀甀攀渀挀椀渀最椀渀漀爀搀攀爀琀漀瘀攀爀椀昀礀琀栀攀攀砀椀猀琀攀渀挀攀漀昀爀攀挀漀洀戀椀渀愀琀椀漀渀⸀

Keyword: hsp70 gene, Paracoccidioides lutzii, Polymerase Chain Reaction

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