Proficiency testing and cross-laboratory method comparison to support standardisation of diatom DNA metabarcoding for freshwater biomonitoring

By

Vasselon, Valentin
Rivera, Sinziana F.
Ács, Éva
Almeida, Salomé F.P.
Andree, Karl B.
Apothéloz-Perret-Gentil, Laure
Bailet, Bonnie
Baricevic, Ana
Beentjes, Kevin K.
Bettig, Juliane
Bouchez, Agnès
Capelli, Camilla
Chardon, Cécile
Duleba, Mónika
Elersek, Tina
Genthon, Clémence
Jablonska, Maša
Jacas, Louis
Kahlert, Maria
Kelly, Martyn G.
Macher, Jan-Niklas
Mauri, Federica
Moletta-Denat, Marina
Mortágua, Andreia
Pawlowski, Jan
Pérez-Burillo, Javier
Pfannkuchen, Martin
Pilgrim, Erik
Pissaridou, Panayiota
Rimet, Frederic
Stanic, Karmen
Tapolczai, Kálmán
Theroux, Susanna
Trobajo, Rosa
Van Der Hoorn, Berry
Vasquez, Marlen I.
Vidal, Marie
Wanless, David
Warren, Jonathan
Zimmermann, Jonas
Paix, Benoît

Type

Article

Date of Publication

2025-1-10

Original Publication

Metabarcoding and metagenomics

Volume

9

Pages

1--36

Subjects

Cross-laboratory experiment , DNA-based approach , ecological status assessment , intercalibration , Standardisation

Contributed by

Pensoft Publishers

Language

English

Published by

Bulgaria Pensoft Publishers 2017-

Identifiers

DOI: https://doi.org/10.3897/mbmg.9.133264

 

Abstract

DNA metabarcoding of benthic diatoms has been successfully applied for biomonitoring at the national scale and can now be considered technically ready for routine application. However, protocols and methods still vary between and within countries, limiting their transferability and the comparability of results. In order to overcome this, routine use of DNA metabarcoding for diatom biomonitoring requires knowledge of the sources of variability introduced by the different steps of the procedure. Here, we examine how elements of routine procedures contribute to variability between European laboratories. A set of four experiments were performed focusing on DNA extraction and PCR amplification steps to evaluate their reproducibility between different laboratories and the variability introduced by different protocols currently applied by the scientific community. Under the guidance of a reference laboratory, 17 participants from 14 countries performed DNA extraction and PCR amplification in parallel, using the same fixed protocol and their own choice of protocol. Experiments were performed by each participant on a set of standardised DNA and biofilm samples (river, lake and mock community) to investigate potential systematic and random errors. Our results revealed the successful transferability of a protocol amongst labs and a highly similar and consistent ecological assessment outcome obtained regardless of the protocols used by each participant. We propose an “all for one but prove them all” strategy, suggesting that distinct protocols can be used within the scientific community, as long as their consistency is be proven by following minimum standard requirements.