Evaluation of viral genome assembly and diversity estimation in deep metagenomes

BMC Genomics

Antonio Alcamí¹  Florent E Angly²  Daniel Aguirre de Cárcer¹ 

[1] Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas (CSIC)–Universidad Autónoma de Madrid, Madrid, Spain;Australian Centre for Ecogenomics, School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, Brisbane, QLD 4072, Australia

关键词: Virome;    Metagenomics;    Diversity;    Assembly;   

Others  :  1092302
DOI  :  10.1186/1471-2164-15-989

 received in 2014-08-24, accepted in 2014-10-30,  发布年份 2014

【 摘 要 】

Background

Viruses have unique properties, small genome and regions of high similarity, whose effects on metagenomic assemblies have not been characterized so far. This study uses diverse in silico simulated viromes to evaluate how extensively genomes can be assembled using different sequencing platforms and assemblers. Further, it investigates the suitability of different methods to estimate viral diversity in metagenomes.

Results

We created in silico metagenomes mimicking various platforms at different sequencing depths. The CLC assembler revealed subpar compared to IDBA_UD and CAMERA , which are metagenomic-specific. Up to a saturation point, Illumina platforms proved more capable of reconstructing large portions of viral genomes compared to 454. Read length was an important factor for limiting chimericity, while scaffolding marginally improved contig length and accuracy. The genome length of the various viruses in the metagenomes did not significantly affect genome reconstruction, but the co-existence of highly similar genomes was detrimental. When evaluating diversity estimation tools, we found that PHACCS results were more accurate than those from CatchAll and clustering, which were both orders of magnitude above expected.

Conclusions

Assemblers designed specifically for the analysis of metagenomes should be used to facilitate the creation of high-quality long contigs. Despite the high coverage possible, scientists should not expect to always obtain complete genomes, because their reconstruction may be hindered by co-existing species bearing highly similar genomic regions. Further development of metagenomics-oriented assemblers may help bypass these limitations in future studies. Meanwhile, the lack of fully reconstructed communities keeps methods to estimate viral diversity relevant. While none of the three methods tested had absolute precision, only PHACCS was deemed suitable for comparative studies.

【 授权许可】

   
2014 Aguirre de Cárcer et al.; licensee BioMed Central Ltd.

【 图 表 】

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