TY - JOUR
T1 - A new duplex PCR assay for the rapid screening of mating-type idiomorphs of pathogenic Sporothrix species
AU - de Carvalho, Jamile Ambrósio
AU - Pinheiro, Breno Gonçalves
AU - Hagen, Ferry
AU - Gonçalves, Sarah Santos
AU - Negroni, Ricardo
AU - Kano, Rui
AU - Bonifaz, Alexandro
AU - de Camargo, Zoilo Pires
AU - Rodrigues, Anderson Messias
N1 - Funding Information:
This research was funded by the Fundação de Amparo à Pesquisa do Estado de São Paulo ( 2017/27265–5 ) http://www.fapesp.br/ , the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq 429594/2018-6 ) http://www.cnpq.br/ , and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES 88887.177846/2018–00 ) https://www.capes.gov.br/ . AMR is a CNPq Research Productivity Fellow (CNPq 304902/2020–9 ).
Funding Information:
This research was funded by the Funda??o de Amparo ? Pesquisa do Estado de S?o Paulo (2017/27265?5) http://www.fapesp.br/, the Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq 429594/2018-6) http://www.cnpq.br/, and the Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES 88887.177846/2018?00) https://www.capes.gov.br/. AMR is a CNPq Research Productivity Fellow (CNPq 304902/2020?9).
Publisher Copyright:
© 2021
PY - 2021/10
Y1 - 2021/10
N2 - 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.
AB - 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.
KW - Duplex PCR
KW - Epidemiology
KW - MAT1-1
KW - MAT1-2
KW - Outbreaks
KW - qPCR
KW - Sporotrichosis
UR - http://www.scopus.com/inward/record.url?scp=85107661710&partnerID=8YFLogxK
U2 - 10.1016/j.funbio.2021.05.005
DO - 10.1016/j.funbio.2021.05.005
M3 - Article
AN - SCOPUS:85107661710
SN - 1878-6146
JO - Fungal Biology
JF - Fungal Biology
ER -