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BENCHMARKING DIFFERENT APPROACHES FOR NOROVIRUS GENOME ASSEMBLY IN METAGENOME SAMPLES
Author
Affilliation
Unit of Genomics and Diabetes. Research Foundation of Valencia University Clinical Hospital- INCLIVA, Valencia, Spain. Spain.
Unit of Genomics and Diabetes. Research Foundation of Valencia University Clinical Hospital- INCLIVA, Valencia, Spain. Spain.
Unit of Genomics and Diabetes. Research Foundation of Valencia University Clinical Hospital- INCLIVA, Valencia, Spain. Spain.
Department of Microbiology, School of Medicine. University of Valencia, Valencia, Spain.
Fundação Oswaldo Cruz. Insituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro. RJ, Brasil. , RJ, Brasil.
Department of Microbiology, School of Medicine. University of Valencia, Valencia, Spain.FUENTES-T
Unit of Genomics and Diabetes. Research Foundation of Valencia University Clinical Hospital- INCLIVA, Valencia, Spain. Spain / 4Spanish Biomedical Research Network in Diabetes and Associated Metabolic Disorders (CIBERDEM), Madrid, Spain.
Department of Microbiology, School of Medicine. University of Valencia, Valencia, Spain.
Department of Microbiology, School of Medicine. University of Valencia, Valencia, Spain / Spanish Biomedical Research Network in Diabetes and Associated Metabolic Disorders (CIBERDEM), Madrid, Spain / Sequecing Multiplex S.L., Valencia, Spain.
Unit of Genomics and Diabetes. Research Foundation of Valencia University Clinical Hospital- INCLIVA, Valencia, Spain. Spain.
Unit of Genomics and Diabetes. Research Foundation of Valencia University Clinical Hospital- INCLIVA, Valencia, Spain. Spain.
Department of Microbiology, School of Medicine. University of Valencia, Valencia, Spain.
Fundação Oswaldo Cruz. Insituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro. RJ, Brasil. , RJ, Brasil.
Department of Microbiology, School of Medicine. University of Valencia, Valencia, Spain.FUENTES-T
Unit of Genomics and Diabetes. Research Foundation of Valencia University Clinical Hospital- INCLIVA, Valencia, Spain. Spain / 4Spanish Biomedical Research Network in Diabetes and Associated Metabolic Disorders (CIBERDEM), Madrid, Spain.
Department of Microbiology, School of Medicine. University of Valencia, Valencia, Spain.
Department of Microbiology, School of Medicine. University of Valencia, Valencia, Spain / Spanish Biomedical Research Network in Diabetes and Associated Metabolic Disorders (CIBERDEM), Madrid, Spain / Sequecing Multiplex S.L., Valencia, Spain.
Abstract
Background: Genome assembly of viruses with high mutation rates, such as Norovirus and other RNA viruses, or
from metagenome samples, poses a challenge for the scientific community due to the coexistence of several viral
quasispecies and strains. Furthermore, there is no standard method for obtaining whole-genome sequences in nonrelated
patients. After polyA RNA isolation and sequencing in eight patients with acute gastroenteritis, we evaluated
two de Bruijn graph assemblers (SPAdes and MEGAHIT), combined with four different and common pre-assembly
strategies, and compared those yielding whole genome Norovirus contigs.
Results: Reference-genome guided strategies with both host and target virus did not present any advantages
compared to the assembly of non-filtered data in the case of SPAdes, and in the case of MEGAHIT, only host
genome filtering presented improvements. MEGAHIT performed better than SPAdes in most samples, reaching
complete genome sequences in most of them for all the strategies employed. Read binning with CD-HIT improved
assembly when paired with different analysis strategies, and more notably in the case of SPAdes.
Conclusions: Not all metagenome assemblies are equal and the choice in the workflow depends on the species
studied and the prior steps to analysis. We may need different approaches even for samples treated equally due to
the presence of high intra host variability. We tested and compared different workflows for the accurate assembly
of Norovirus genomes and established their assembly capacities for this purpose.
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