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⸀