Abstract
Paracoccidioidomycosis (PCM) is a mycotic disease caused by the Paracoccidioides species, a group of thermally dimorphic fungi that grow in mycelial form at 25 °C and as budding yeasts when cultured at 37 °C or when parasitizing the host tissues. PCM occurs in a large area of Latin America, and the most critical regions of endemicity are in Brazil, Colombia, and Venezuela. The clinical diagnosis of PCM needs to be confirmed through laboratory tests. Although classical laboratory techniques provide valuable information due to the presence of pathognomonic forms of Paracoccidioides spp., nucleic acid-based diagnostics gradually are replacing or complementing culture-based, biochemical, and immunological assays in routine microbiology laboratory practice. Recently, taxonomic changes driven by whole-genomic sequencing of Paracoccidioides have highlighted the need to recognize species boundaries, which could better ascertain Paracoccidioides taxonomy. In this scenario, classical laboratory techniques do not have significant discriminatory power over cryptic agents. On the other hand, several PCR-based methods can detect polymorphisms in Paracoccidioides DNA and thus support species identification. This review is focused on the recent achievements in molecular diagnostics of paracoccidioidomycosis, including the main advantages and pitfalls related to each technique. We discuss these breakthroughs in light of taxonomic changes in the Paracoccidioides genus.
Highlights
Discipline of Cellular Biology, Federal University of São Paulo (UNIFESP), São Paulo 04023062, Brazil; Laboratory of Mycology/Research, Faculty of Medicine, Federal University of Mato Grosso, Cuiabá, Federal University of Mato Grosso, Júlio Muller University Hospital, Mato Grosso 78048902, Brazil
The results suggested that serum samples were not suited for polymerase chain reaction (PCR) diagnosis of either species
Transposable elements (Trems) are estimated to comprise approximately 16% of the P. lutzii genome and 9% of P. brasiliensis, with the possibility to distinguish between cryptic species [117]
Summary
The classical PCM diagnosis combines clinical evaluation and additional laboratory investigations, including routine culture-based, biochemical, and immunological assays in microbiology laboratories (Figure 3). Immunodiagnostics include techniques such as counterimmunoelectrophoresis reaction (CIE) [41], enzyme-linked immunosorbent assay (ELISA) [42], latex agglutination assay [43], and immunoblotting [44], which are available from different reference services in Brazil (Figure 3) These tests usually employ a 43 kDa glycoprotein (GP43) as the P. brasiliensis complex’s primary antigen to detect circulating antibodies and have sensitivity between 85% and 100% [38]. As an alternative to classical laboratory diagnosis, molecular assays can be used for Paracoccidioides spp. aiming at different strategies, such as detection and identification In this scenario, molecular-based assays provide the fastest and most accurate results about the infection, reaching species level more efficiently [21]. The literature search criteria utilized for the preparation of this article are identified in the Supplementary Material (Table S1)
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