Deep Impact of Random Amplification and Library Construction Methods on Viral Metagenomics Results.

ABSTRACT: Clinical metagenomics is a broad-range agnostic detection method of pathogens, including novel microorganisms. A major limit is the low pathogen load compared to the high background of host nucleic acids. To overcome this issue, several solutions exist, such as applying a very high depth of sequencing, or performing a relative enrichment of viral genomes associated with capsids. At the end, the quantity of total nucleic acids is often below the concentrations recommended by the manufacturers of library kits, which necessitates to random amplify nucleic acids. Using a pool of 26 viruses representative of viral diversity, we observed a deep impact of the nature of sample (total nucleic acids versus RNA only), the reverse transcription, the random amplification and library construction method

SUBMITTER: Regnault B 

PROVIDER: S-EPMC7915491 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature