A new duplex PCR assay for the rapid screening of mating-type idiomorphs of pathogenic Sporothrix species

Author(s)

de Carvalho, Jamile Ambrosio ^{[1, 2]} ; Pinheiro, Breno Goncalves ^[1] ; Hagen, Ferry ^{[3, 4, 5]} ; Goncalves, Sarah Santos ^[6] ; Negroni, Ricardo ^[7] ; Kano, Rui ^[8] ; Bonifaz, Alexandro ^[9] ; de Camargo, Zoilo Pires ^{[1, 2]} ; Rodrigues, Anderson Messias ^{[1, 2]}

Affiliation

^[1] Fed Univ Sao Paulo UNIFESP, Dept Microbiol Immunol & Parasitol, Lab Emerging Fungal Pathogens, Discipline Cellular Biol, BR-04023062 Sao Paulo - Brazil

^[2] Fed Univ Sao Paulo UNIFESP, Dept Med, Discipline Infect Dis, BR-04023062 Sao Paulo - Brazil

^[3] Jining 1 Peoples Hosp, Lab Med Mycol, Jining, Shandong - Peoples R China

^[4] Westerdijk Fungal Biodivers Inst, Dept Med Mycol, Uppsalalaan 8, NL-3584 CT Utrecht - Netherlands

^[5] Univ Med Ctr Utrecht, Dept Med Microbiol, Heidelberglaan 100, NL-3584 CX Utrecht - Netherlands

^[6] Fed Univ Espirito Santo UFES, Ctr Res Med Mycol, Infect Dis Postgrad Program, Vitoria, ES - Brazil

^[7] Infect Dis Hosp FJ Muniz, Reference Ctr Mycol Buenos Aires City, Mycol Unit, Buenos Aires, DF - Argentina

^[8] Nihon Univ, Dept Vet Dermatol, Coll Bioresource Sci, Fujisawa, Kanagawa - Japan

^[9] Hosp Gen Mexico Dr Eduardo Liceaga, Mycol Dept, Dermatol Serv, Mexico City, DF - Mexico

• Document type: Journal article
• Source: FUNGAL BIOLOGY; v. 125, n. 10, p. 834-843, OCT 2021.
• Web of Science Citations: 1

Abstract

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 (alpha-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; chi(2) = 10.286, P = 0.0013) opposing the high prevalence MAT1-1 in the Rio Grande do Sul (10:1 ratio; chi(2) = 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. (C) 2021 British Mycological Society. Published by Elsevier Ltd. All rights reserved. (AU)

• FAPESP's process: 17/27265-5 - Molecular epidemiology and genomic perspectives on the evolution and spread of emerging fungal pathogens
• Grantee: Anderson Messias Rodrigues
• Support Opportunities: Research Grants - Young Investigators Grants