Analysis of genetic diversity in aspergillosis agents using AFLP (Amplified Fragment Length Polymorphism) and microsatellites markers

• Grant number: 21/02327-3
• Support Opportunities: Scholarships in Brazil - Master
• Effective date (Start): April 01, 2021
• Effective date (End): March 31, 2023
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: Anderson Messias Rodrigues
• Grantee: Úrsula dos Santos Lopez
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated research grant: 17/27265-5 - Molecular epidemiology and genomic perspectives on the evolution and spread of emerging fungal pathogens, AP.JP

Abstract

The major agent of aspergillosis is Aspergillus fumigatus (85%-90%), an omnipresent fungus that causes a variety of clinical lung syndromes, ranging from aspergilloma in patients with pulmonary cavities to chronic necrotizing aspergillosis in slightly immunocompromised people or those with chronic lung disease. In this scenario, invasive pulmonary aspergillosis is highlighted, which is a severe and commonly fatal disease observed in immunocompromised patients. There is a growing list of emerging agents in Aspergillus that includes A. flavus, A. terreus, A. niger, and A. nidulans. This situation is problematic due to the absence of epidemiological studies associated with the difficulty of diagnosis and the emergence of resistance to azoles used as treatment. Thus, this work aims to evaluate the technique of AFLP as an epidemiological tool of molecular typing to investigate the genetic diversity and population structure in agents of human aspergillosis in Brazil. Population differentiation in Aspergillus will be investigated, and gene flow patterns will be estimated. The existence of initial stages of speciation and the probable occurrence of cryptic species will be evaluated together with the results of nuclear loci sequencing (MLSA). AFLP performance will be compared with other standard molecular tools such as microsatellite markers (STRAf 2A, 2B, 2C, 3A, 3B, 3C, 4A, 4B, and 4C). We will evaluate the presence and distribution of sexual idiomorphs MAT 1-1 and 1-2 among the isolates inserted in this population. Finally, the MLSA technique will be used to confirm the data obtained from AFLP markers. The data generated in this project will contribute to the knowledge of the epidemiology of the fungus and will serve as a support for further research, aiming at the elaboration of strategies for disease control. (AU)