@article{fdi:010065356,
  title = {{H}ost-associated metagenomics : a guide to generating infectious {RNA} viromes},
  author = {{T}emmam, {S}. and {M}onteil-{B}ouchard, {S}. and {R}obert, {C}. and {P}ascalis, {H}erv{\'e} and {M}ichelle, {C}. and {J}ardot, {P}. and {C}harrel, {R}. and {R}aoult, {D}idier and {D}esnues, {C}.},
  editor = {},
  language = {{ENG}},
  abstract = {{B}ackground {M}etagenomic analyses have been widely used in the last decade to describe viral communities in various environments or to identify the etiology of human, animal, and plant pathologies. {H}ere, we present a simple and standardized protocol that allows for the purification and sequencing of {RNA} viromes from complex biological samples with an important reduction of host {DNA} and {RNA} contaminants, while preserving the infectivity of viral particles. {P}rincipal {F}indings {W}e evaluated different viral purification steps, random reverse transcriptions and sequence-independent amplifications of a pool of representative {RNA} viruses. {V}iruses remained infectious after the purification process. {W}e then validated the protocol by sequencing the {RNA} virome of human body lice engorged in vitro with artificially contaminated human blood. {T}he full genomes of the most abundant viruses absorbed by the lice during the blood meal were successfully sequenced. {I}nterestingly, random amplifications differed in the genome coverage of segmented {RNA} viruses. {M}oreover, the majority of reads were taxonomically identified, and only 7-15% of all reads were classified as "unknown", depending on the random amplification method. {C}onclusion {T}he protocol reported here could easily be applied to generate {RNA} viral metagenomes from complex biological samples of different origins. {O}ur protocol allows further virological characterizations of the described viral communities because it preserves the infectivity of viral particles and allows for the isolation of viruses.},
  keywords = {},
  booktitle = {},
  journal = {{P}los {O}ne},
  volume = {10},
  numero = {10},
  pages = {e0139810 [21 p.]},
  ISSN = {1932-6203},
  year = {2015},
  DOI = {10.1371/journal.pone.0139810},
  URL = {https://www.documentation.ird.fr/hor/fdi:010065356},
}