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How quantitative is metabarcoding: A meta-analytical approach.

Philip D Lamb1, Ewan Hunter2,3, John K Pinnegar2,3

  • 1School of Biological Sciences, University of East Anglia, Norwich, UK.

Molecular Ecology
|November 9, 2018
PubMed
Summary
This summary is machine-generated.

Metabarcoding shows a weak quantitative link between species biomass and sequence reads, with high uncertainty. More research is needed for reliable quantitative ecological studies using this method.

Keywords:
biomasshigh-throughput sequencingmeta-analysismetabarcodingnext-generation sequencing

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Area of Science:

  • Ecology
  • Molecular Biology
  • Bioinformatics

Background:

  • Metabarcoding is widely used in ecology for taxonomic assignment, dietary analysis, and environmental DNA (eDNA) studies.
  • Despite a decade of application, there is a lack of consensus on the quantitative accuracy of metabarcoding, specifically how read proportions relate to original species proportions.

Purpose of the Study:

  • To systematically review and meta-analyze existing research to quantify the current understanding of metabarcoding's quantitative performance.
  • To assess the relationship between species biomass and the number of sequence reads generated.

Main Methods:

  • Conducted a structured review of published metabarcoding studies.
  • Performed a meta-analysis to statistically evaluate the relationship between biomass and sequence data.
  • Tested potential moderators including sequencing platform, species number, and DNA source to explain variance.

Main Results:

  • A weak, yet statistically significant, quantitative relationship was found between species biomass and sequence reads (slope = 0.52 ± 0.34, p < 0.01).
  • Significant uncertainty surrounds this relationship.
  • No tested moderators (sequencing platform, species number, DNA source) could explain the observed variance in quantitative performance.

Conclusions:

  • Current understanding of factors influencing metabarcoding's quantitative accuracy is limited.
  • Further research is essential to establish confidence in using metabarcoding for quantitative ecological applications.
  • The use of mock communities is recommended to directly assess the quantitative capabilities of metabarcoding studies until its reliability is better established.