Proficiency testing and cross-laboratory method comparison to support standardisation of diatom DNA metabarcoding for freshwater biomonitoring
Abstrak
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.
Topik & Kata Kunci
Penulis (41)
Valentin Vasselon
Sinziana F. Rivera
Éva Ács
Salomé Baja Almeida
Karl B. Andree
Laure Apothéloz-Perret-Gentil
Bonnie Bailet
Ana Baričević
Kevin K. Beentjes
Juliane Bettig
Agnès Bouchez
Camilla Capelli
Cécile Chardon
Mónika Duleba
Tina Elersek
Clémence Genthon
Maša Jablonska
Louis Jacas
Maria Kahlert
Martyn G. Kelly
Jan-Niklas Macher
Federica Mauri
Marina Moletta-Denat
Andreia Mortágua
Jan Pawlowski
Javier Pérez-Burillo
Martin Pfannkuchen
Erik Pilgrim
Panayiota Pissaridou
Frédéric Rimet
Karmen Stanic
Kálmán Tapolczai
Susanna Theroux
Rosa Trobajo
Berry Van der Hoorn
Marlen I. Vasquez
Marie Vidal
David Wanless
Jonathan Warren
Jonas Zimmermann
Benoît Paix
Format Sitasi
Akses Cepat
- Tahun Terbit
- 2025
- Sumber Database
- DOAJ
- DOI
- 10.3897/mbmg.9.133264
- Akses
- Open Access ✓