Macroporous epoxy-based monoliths for rapid quantification of Pseudomonas aeruginosa by adsorption elution method optimi
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RESEARCH PAPER
Macroporous epoxy-based monoliths for rapid quantification of Pseudomonas aeruginosa by adsorption elution method optimized for qPCR Lisa Göpfert 1 & Julia Klüpfel 1 & Charlotte Heinritz 1 & Martin Elsner 1 & Michael Seidel 1 Received: 1 July 2020 / Revised: 11 September 2020 / Accepted: 16 September 2020 / Published online: 3 October 2020 # The Author(s) 2020
Abstract Pseudomonas aeruginosa contaminations in tap water systems have caused severe health problems in both hospital and household settings. To ensure fast and reliable detection, culture-independent methods are recommendable. However, the typically low cell number in water samples requires sample enrichment prior to analysis. Therefore, we developed and optimized an adsorption elution method using monolithic adsorption filtration and subsequent centrifugal ultrafiltration that can be combined with culture-independent detection methods. The principle of adsorption of Pseudomonas aeruginosa by hydrophobic and ionic interactions was studied in modified epoxy-based monoliths. Optimized conditions (5-L initial sample volume at pH 3 filtered for 30 min through hydrolyzed monoliths (MAF-OH) and eluted with beef extract glycine buffer at pH 9.5) achieved a recovery of 67.1 ± 1.2% and a concentration factor of 103. For the first time, we therefore present a culture-independent approach for rapid enrichment and subsequent molecular biological quantification of P. aeruginosa by qPCR from tap water samples by monolithic adsorption filtration. The total enrichment and quantification process takes 4 h. This work further stresses the versatility of the monolithic adsorption filtration and its possibilities as a concentration tool for culture-independent analytics of pathogenic bacteria in the environment. Keywords Monolith filtration . Pseudomonas aeruginosa . Culture-independent . Adsorption elution
Introduction Providing safe drinking water — or lowering risk for workers in contact with technical water — nowadays requires very sensitive analytical techniques to detect and rapidly quantify pathogenic bacteria. Therefore, it is necessary to concentrate bacteria from large water sample volumes prior to subsequent analysis by cultivation-independent methods. Here, traditional sterile filtration has its disadvantages as the filters tend to clog quickly due to matrix components when large water sample Lisa Göpfert and Julia Klüpfel contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00216-020-02956-3) contains supplementary material, which is available to authorized users. * Michael Seidel [email protected] 1
Institute of Hydrochemistry, Chair of Analytical Chemistry and Water Chemistry, Technical University of Munich, Marchioninistr. 17, 81377 Munich, Germany
volumes are processed. Monolith-based filtration offers an attractive alternative. Monoliths are homogeneous stationary phases with high porosity. They can be comprised of organic or inorganic material, most often silica
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