Genotyping ofParacoccidioides brasiliensisdirectly from paraffin embedded tissue

Abstract

Based on polymorphisms of the gp43 precursor gene, genotyping of Paracoccidioides brasiliensis was achieved in 6 out of 10 paraffin-embedded tissue samples by modifying a nested PCR procedure used in the diagnosis of the fungal infection. Nine of the samples originated in Brazil. Three sequences were identical to a previously published consensus sequence identifying the P. brasiliensis isolates as members of the formerly named species 1. In contrast, two sequences revealed substitutions identical to isolates of the phylogenetic species 2. Applying the method to a lung biopsy from a proven case of paracoccidioidomycosis diagnosed in Austria, the gp43 sequence revealed substitutions so far only described in five strains originating from Venezuela. Combining travel history and result of this new method, the most probable country of origin of the infections could be identified. Since endemic mycosis are often diagnosed by histopathology only, our method could help to extend epidemiological studies of paracoccidioidomycosis.

Clinical features and genetic background of the sympatric species Paracoccidioides brasiliensis and Paracoccidioides americana

“…americana and P . brasiliensis samples from Southeast Brazil (Rio de Janeiro, São Paulo and Minas Gerais) were retrieved for counts [12, 14, 36–42]. These three states are located in the Southeastern part of Brazil, a hot spot of PCM, and contain the majority of currently genotyped strains of Paracoccidioides .…”

Molecular approaches for eco-epidemiological studies of Paracoccidioides brasiliensis

“…Since gp43 is a highly polymorphic gene that clearly separates the four cryptic species of P. brasiliensis (Matute et al 2006a, Carrero et al 2008, Teixeira 2008, PCR based on this genomic region is now indicated not only for pathogen detection, but also for genotyping of clinical samples. By using this strategy, Ricci et al (2008) evaluated and distinguished between S1-PS2 from paraffin-embedded tissue.…”

Case Report: Paracoccidioidomycosis in Solid Organ Transplantation: Disseminated Disease in a Liver Recipient and Literature Review

“…Molecular identification was conducted by sequencing the gp43 gene of P. brasiliensis extracted from paraffin-embedded tissue using the para I and para IV primers, as described in the literature. 10 Disseminated disease was confirmed with multiple cutaneous abscesses, lung injury, and significant impairment of adrenal, reticuloendothelial, osteoarticular, and integumentary systems, which were noted on thoracic and abdominal computed tomography ( Figure 2). The patient tested negative for HIV-1 and -2 and hepatitis B and C. Amphotericin B complex lipid induction therapy was initiated at 200 mg/day for 20 days, followed by maintenance treatment with TMP-SMX 800/160 mg, twice daily.…”