Autor: |
Moreno KMF; Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte 31270-901, Brazil., de Andrade VA; Eduardo de Menezes Hospital, Belo Horizonte 30622-020, Brazil., de Melo Iani FC; Central Public Health Laboratory of the State of Minas Gerais, Ezequiel Dias Foundation, Belo Horizonte 30510-010, Brazil., Fonseca V; Department of Exact and Earth Sciences, University of the State of Bahia, Salvador 41150-000, Brazil., Lima MT; Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte 31270-901, Brazil.; René Rachou Institute, Oswaldo Cruz Foundation, Belo Horizonte 30190-002, Brazil., de Castro Barbosa E; René Rachou Institute, Oswaldo Cruz Foundation, Belo Horizonte 30190-002, Brazil., Tomé LMR; René Rachou Institute, Oswaldo Cruz Foundation, Belo Horizonte 30190-002, Brazil., Guimarães NR; René Rachou Institute, Oswaldo Cruz Foundation, Belo Horizonte 30190-002, Brazil., Fritsch HM; Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte 31270-901, Brazil.; Morphogenesis and Antigenicity of HIV and Hepatitis Viruses, University of Tours, 37032 Tours, France., Adelino T; Central Public Health Laboratory of the State of Minas Gerais, Ezequiel Dias Foundation, Belo Horizonte 30510-010, Brazil., Oliveira Fereguetti T; Eduardo de Menezes Hospital, Belo Horizonte 30622-020, Brazil., Aspahan MC; Eduardo de Menezes Hospital, Belo Horizonte 30622-020, Brazil., Gamarano Barros T; Eduardo de Menezes Hospital, Belo Horizonte 30622-020, Brazil., Alcantara LCJ; René Rachou Institute, Oswaldo Cruz Foundation, Belo Horizonte 30190-002, Brazil., Giovanetti M; Department of Sciences and Technologies for Sustainable Development and One Health, Università Campus Bio-Medico di Roma, 00128 Rome, Italy.; Oswaldo Cruz Foundation, Rio de Janeiro 21040-900, Brazil. |
Abstrakt: |
Acute febrile illness (AFI) and severe neurological disorders (SNDs) often present diagnostic challenges due to their potential origins from a wide range of infectious agents. Nanopore metagenomics is emerging as a powerful tool for identifying the microorganisms potentially responsible for these undiagnosed clinical cases. In this study, we aim to shed light on the etiological agents underlying AFI and SND cases that conventional diagnostic methods have not been able to fully elucidate. Our approach involved analyzing samples from fourteen hospitalized patients using a comprehensive nanopore metagenomic approach. This process included RNA extraction and enrichment using the SMART-9N protocol, followed by nanopore sequencing. Subsequent steps involved quality control, host DNA/cDNA removal, de novo genome assembly, and taxonomic classification. Our findings in AFI cases revealed a spectrum of disease-associated microbes, including Escherichia coli , Streptococcus sp., Human Immunodeficiency Virus 1 (Subtype B), and Human Pegivirus. Similarly, SND cases revealed the presence of pathogens such as Escherichia coli , Clostridium sp., and Dengue virus type 2 (Genotype-II lineage). This study employed a metagenomic analysis method, demonstrating its efficiency and adaptability in pathogen identification. Our investigation successfully identified pathogens likely associated with AFI and SNDs, underscoring the feasibility of retrieving near-complete genomes from RNA viruses. These findings offer promising prospects for advancing our understanding and control of infectious diseases, by facilitating detailed genomic analysis which is critical for developing targeted interventions and therapeutic strategies. |