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• W2216893304 endingPage "e0004190" @default.
• W2216893304 startingPage "e0004190" @default.
• W2216893304 abstract "Sporotrichosis is a chronic (sub)cutaneous infection caused by thermodimorphic fungi in the order, Ophiostomatales. These fungi are characterized by major differences in routes of transmission, host predilections, species virulence, and susceptibilities to antifungals. Sporothrix species emerge in the form of outbreaks. Large zoonoses and sapronoses are ongoing in Brazil and China, respectively. Current diagnostic methods based on morphology and physiology are inaccurate due to closely related phenotypes with overlapping components between pathogenic and non-pathogenic Sporothrix. There is a critical need for new diagnostic tools that are specific, sensitive, and cost-effective.We developed a panel of novel markers, based on calmodulin (CAL) gene sequences, for the large-scale diagnosis and epidemiology of clinically relevant members of the Sporothrix genus, and its relative, Ophiostoma. We identified specific PCR-based markers for S. brasiliensis, S. schenckii, S. globosa, S. mexicana, S. pallida, and O. stenoceras. We employed a murine model of disseminated sporotrichosis to optimize a PCR assay for detecting Sporothrix in clinical specimens.Primer-BLAST searches revealed candidate sequences that were conserved within a single species. Species-specific primers showed no significant homology with human, mouse, or microorganisms outside the Sporothrix genus. The detection limit was 10-100 fg of DNA in a single round of PCR for identifying S. brasiliensis, S. schenckii, S. globosa, S. mexicana, and S. pallida. A simple, direct PCR assay, with conidia as a source of DNA, was effective for rapid, low-cost genotyping. Samples from a murine model of disseminated sporotrichosis confirmed the feasibility of detecting S. brasiliensis and S. schenckii DNA in spleen, liver, lungs, heart, brain, kidney, tail, and feces of infected animals.This PCR-based method could successfully detect and identify a single species in samples from cultures and from clinical specimens. The method proved to be simple, high throughput, sensitive, and accurate for diagnosing sporotrichosis." @default.
• W2216893304 created "2016-06-24" @default.
• W2216893304 creator A5013215405 @default.
• W2216893304 creator A5062912750 @default.
• W2216893304 creator A5085601234 @default.
• W2216893304 date "2015-12-01" @default.
• W2216893304 modified "2023-10-16" @default.
• W2216893304 title "Molecular Diagnosis of Pathogenic Sporothrix Species" @default.
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• W2216893304 doi "https://doi.org/10.1371/journal.pntd.0004190" @default.
• W2216893304 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4666615" @default.
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• W2216893304 hasPublicationYear "2015" @default.
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