| Congresso Brasileiro de Microbiologia 2023 | Resumo: 1224-1 | ||||
Resumo:Sporotrichosis is a cutaneous or subcutaneous mycosis caused by thermodimorphic Sporothrix species. Of particular concern is the emergence of cat-transmitted sporotrichosis associated with Sporothrix brasiliensis, which has seen an alarming rise in cases in Brazil. Infected cats act as sources of the fungal propagules responsible for human infections, posing a potential public health threat. Understanding the dynamics of Sporothrix transmission in the context of the feline population is crucial for implementing effective control measures and raising awareness about the risks associated with close contact with infected animals. Recently, molecular techniques have been used to describe the genetic diversity of Sporothrix spp., particularly in S. brasiliensis and S. schenckii. Choosing appropriate methods with good cost-benefit and effectiveness is crucial for optimizing the genetic diversity analysis process. This study aimed to compare two molecular techniques, AFLP (Amplified fragment length polymorphism) and SSR (Simple sequence repeats), to determine the most suitable method for analyzing genetic diversity in medically relevant Sporothrix species. To achieve this, 30 samples of Sporothrix spp. from the clinical clade (S. brasiliensis n=10, S. schenckii n=10, and S. globosa n=10) were tested using three combinations of AFLP markers (#3 EcoRI-GA/MseI-TT, #5 EcoRI-GA/MseI-AG, and #6 EcoRI-TA/MseI-AA) and 15 SSR markers. The discriminatory power of each technique in studying genetic variability was assessed. The data obtained were analyzed using BioNumerics v.7.6, and dendrograms (Jaccard Distance, UPGMA) were constructed. The dendrograms displayed well-supported branches with global cophenetic values greater than 98% for AFLP and 99% for SSR, indicating the reliability of both techniques. Genetic diversity was calculated for AFLP and SSR, with values of polymorphism information content (PIC) ranging from 0.1144 to 0.2879, marker index (MI) from 0.0026 to 0.1208, effective multiplex ratio (E) from 1.0000 to 63.2333, discriminating power (D) from 0.0968 to 0.9493, expected heterozygosity (H) from 0.1208 to 0.3487, and mean heterozygosity (Havp) from 0.00004 to 0.12085. These results revealed the utility of both AFLP and SSR in discriminating among different Sporothrix spp. Principal component analysis (PCA) plots were generated, showing a cumulative percentage of 54.5% for AFLP and 59.4% for the set of SSR. The congruence between the experiments was determined to be 80.98%, with a very good agreement observed between AFLP and SSR (Kappa=1.0; 95% CI=1.0–1.0), further supporting the effectiveness of these tests for genotyping Sporothrix spp. Although both AFLP and SSR were found to estimate genetic diversity among Sporothrix spp. other practical factors such as costs, protocol execution time, difficulty level, and labor errors during the protocol execution must be considered. Thus, SSR proves to be a less expensive, faster, and more straightforward technique, yielding essential results for the epidemiological investigation of sporotrichosis. We recommend using at least one combination of AFLP (#5 EcoRI-GA/MseI-AG) and the complete panel of 15 SSR markers for more accurate diversity results. The study of genetic diversity has emerged as a valuable tool in epidemiological studies and will play a crucial role in improving public policy strategies to raise awareness about the spread of Sporothrix and to control sporotrichosis effectively. Palavras-chave: AFLP, genetic diversity, Sporothrix, sporotrichosis, SSR Agência de fomento:CAPES (88887.159096/2017-00), CNPq (433276/2018-5), FAPESP (2017/27265-5) | |||||