A new duplex PCR assay for the rapid screening of mating-type idiomorphs of pathogenic Sporothrix species
| Autor: | Ferry Hagen, Breno Gonçalves Pinheiro, Sarah Santos Gonçalves, Jamile Ambrósio de Carvalho, Zoilo Pires de Camargo, Anderson Messias Rodrigues, Alexandro Bonifaz, Ricardo Negroni, Rui Kano |
|---|---|
| Přispěvatelé: | Westerdijk Fungal Biodiversity Institute, Westerdijk Fungal Biodiversity Institute - Medical Mycology |
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
Mating type Epidemiology Locus (genetics) Polymerase Chain Reaction 01 natural sciences 03 medical and health sciences Genetics medicine Animals Sporothrix schenckii Mating Ecology Evolution Behavior and Systematics 030304 developmental biology 0303 health sciences biology Sporotrichosis Sporothrix Duplex PCR Outbreaks MAT1-2 Outbreak MAT1-1 Amplicon biology.organism_classification medicine.disease qPCR Infectious Diseases Brazil 010606 plant biology & botany |
| Zdroj: | Fungal Biology. Elsevier B.V. |
| ISSN: | 1878-6146 |
| DOI: | 10.1016/j.funbio.2021.05.005 |
| Popis: | Sporothrix schenckii and allied species are thermodimorphic fungi widely distributed in nature which causes human and animal sporotrichosis, the most common subcutaneous mycosis globally. Sporotrichosis is acquired after a traumatic inoculation of soil or plant material contaminated with Sporothrix propagules or through bites and scratches from diseased cats. In Ascomycota, the master regulators of sex are MAT genes that lie in a single mating-type locus, in Sporothrix these are determined by two nonhomologous alleles, MAT1-1 and MAT1-2. We assessed the whole-genome sequences of medically relevant Sporothrix to develop a single-tube duplex PCR assay to screen S. brasiliensis, S. schenckii, S. globosa, and S. luriei idiomorphs (MAT1-1 or MAT1-2) and understand the distribution and incidence of mating-type strains from natural populations. Using our duplex PCR assay, a 673 bp amplicon (α-box protein) was consistently amplified from all MAT1-1 isolates, while a 291 bp fragment was only amplified from the isolates harboring MAT1-2 (HMG box). Molecular evidence suggests heterothallism (self-sterility) as the unique mating strategy among the species evaluated. The mating-type identity of 93 isolates revealed a nearly equal distribution (1:1 ratio) of mating type alleles within species but deviating between different outbreak areas. Remarkably, for S. brasiliensis in Rio de Janeiro, we report an overwhelming occurrence of MAT1-2 (1:13 ratio; χ2 = 10.286, P = 0.0013) opposing the high prevalence MAT1-1 in the Rio Grande do Sul (10:1 ratio; χ2 = 7.364, P = 0.0067). Therefore, the population structure of Sporothrix species refers from paucity to regular cycles of sexual recombination in most of the studied regions. Our PCR-based mating-type diagnostic assay is proposed here as an important marker to track the geographical expansion during the long-lasting outbreak of cat-transmitted sporotrichosis driven by S. brasiliensis. |
| Databáze: | OpenAIRE |
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