Quality control of microbiota metagenomics by k-mer analysis

Research output: Contribution to journalJournal articleResearchpeer-review

  • Florian Plaza Onate
  • Jean Michel Batto
  • Catherine Juste
  • Jehane Fadlallah
  • Fougeroux, Cyrielle Elyette
  • Doriane Gouas
  • Nicolas Pons
  • Sean Kennedy
  • Florence Levenez
  • Joel Dore
  • S. Dusko Ehrlich
  • Guy Gorochov
  • Martin Larsen

Background: The biological and clinical consequences of the tight interactions between host and microbiota are rapidly being unraveled by next generation sequencing technologies and sophisticated bioinformatics, also referred to as microbiota metagenomics. The recent success of metagenomics has created a demand to rapidly apply the technology to large case-control cohort studies and to studies of microbiota from various habitats, including habitats relatively poor in microbes. It is therefore of foremost importance to enable a robust and rapid quality assessment of metagenomic data from samples that challenge present technological limits (sample numbers and size). Here we demonstrate that the distribution of overlapping k-mers of metagenome sequence data predicts sequence quality as defined by gene distribution and efficiency of sequence mapping to a reference gene catalogue. Results: We used serial dilutions of gut microbiota metagenomic datasets to generate well-defined high to low quality metagenomes. We also analyzed a collection of 52 microbiota-derived metagenomes. We demonstrate that k-mer distributions of metagenomic sequence data identify sequence contaminations, such as sequences derived from "empty" ligation products. Of note, k-mer distributions were also able to predict the frequency of sequences mapping to a reference gene catalogue not only for the well-defined serial dilution datasets, but also for 52 human gut microbiota derived metagenomic datasets. Conclusions: We propose that k-mer analysis of raw metagenome sequence reads should be implemented as a first quality assessment prior to more extensive bioinformatics analysis, such as sequence filtering and gene mapping. With the rising demand for metagenomic analysis of microbiota it is crucial to provide tools for rapid and efficient decision making. This will eventually lead to a faster turn-around time, improved analytical quality including sample quality metrics and a significant cost reduction. Finally, improved quality assessment will have a major impact on the robustness of biological and clinical conclusions drawn from metagenomic studies.

Original languageEnglish
Article number183
JournalBMC Genomics
Volume16
Issue number1
ISSN1471-2164
DOIs
Publication statusPublished - 14 Mar 2015

Bibliographical note

Funding Information:
The authors acknowledge the funding agencies and the volunteers providing samples for the study. The study was funded by INSERM, the University Pierre et Marie Curie ËMERGENCE” program, Fondation pour l’Aide a la Recherche sur la Sclerose En Plaques (ARSEP), ARTHRITIS Fondation COURTIN and Agence nationale de la recherché (ANR). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Publisher Copyright:
© Plaza Onate et al.

    Research areas

  • Metagenomics, Next generation sequencing, Quality control, Sample size limits, Sampling bias

ID: 339849165