Time to act-assessing variations in qPCR analyses in biological nitrogen removal with examples from partial nitritation/anammox systems.

Autor: Agrawal S; Department of Civil and Environmental Engineering Sciences, Institute IWAR, Chair of Wastewater Engineering, Technical University of Darmstadt, Franziska-Braun-Straße 7, 64287 Darmstadt, Germany. Electronic address: s.agrawal@iwar.tu-darmstadt.de., Weissbrodt DG; Department of Biotechnology, Delft University of Technology, Delft, the Netherlands., Annavajhala M; Department of Earth and Environmental Engineering, Columbia University, New York, USA., Jensen MM; Department of Environmental Engineering, Microbial Ecology & Technology Laboratory, Technical University of Denmark, Bygningtorvet, Bldg 115, DK-2800, Lyngby, Denmark., Arroyo JMC; Center of Microbial Ecology and Technology, Ghent University, Coupure Links 653, 9000 Ghent, Belgium., Wells G; Department of Civil and Environmental Engineering, Northwestern University, 2145 Sheridan Road, Evanston, IL, USA., Chandran K; Department of Earth and Environmental Engineering, Columbia University, New York, USA., Vlaeminck SE; Department of Bioscience Engineering, Research Group of Sustainable Energy, Air and Water Technology, University of Antwerp, Groenenborgerlaan 171, 2020 Antwerpen, Belgium., Terada A; Institute of Global Innovation Research and Department of Chemical Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka, Koganei, Tokyo 184-8588, Japan., Smets BF; Department of Environmental Engineering, Microbial Ecology & Technology Laboratory, Technical University of Denmark, Bygningtorvet, Bldg 115, DK-2800, Lyngby, Denmark., Lackner S; Department of Civil and Environmental Engineering Sciences, Institute IWAR, Chair of Wastewater Engineering, Technical University of Darmstadt, Franziska-Braun-Straße 7, 64287 Darmstadt, Germany.
Jazyk: angličtina
Zdroj: Water research [Water Res] 2021 Feb 15; Vol. 190, pp. 116604. Date of Electronic Publication: 2020 Nov 04.
DOI: 10.1016/j.watres.2020.116604
Abstrakt: Quantitative PCR (qPCR) is broadly used as the gold standard to quantify microbial community fractions in environmental microbiology and biotechnology. Benchmarking efforts to ensure the comparability of qPCR data for environmental bioprocesses are still scarce. Also, for partial nitritation/anammox (PN/A) systems systematic investigations are still missing, rendering meta-analysis of reported trends and generic insights potentially precarious. We report a baseline investigation of the variability of qPCR-based analyses for microbial communities applied to PN/A systems. Round-robin testing was performed for three PN/A biomass samples in six laboratories, using the respective in-house DNA extraction and qPCR protocols. The concentration of extracted DNA was significantly different between labs, ranged between 2.7 and 328 ng mg -1 wet biomass. The variability among the qPCR abundance data of different labs was very high (1-7 log fold) but differed for different target microbial guilds. DNA extraction caused maximum variation (3-7 log fold), followed by the primers (1-3 log fold). These insights will guide environmental scientists and engineers as well as treatment plant operators in the interpretation of qPCR data.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2020. Published by Elsevier Ltd.)
Databáze: MEDLINE
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