Macroporous epoxy-based monoliths for rapid quantification of Pseudomonas aeruginosa by adsorption elution method optimized for qPCR

• Published in: Analytical and bioanalytical chemistry 2020-11, Vol.412 (29), p.8185-8195
• Main Authors: Göpfert, Lisa, Klüpfel, Julia, Heinritz, Charlotte, Elsner, Martin, Seidel, Michael
• Format: Article
• Language: English
• Subjects: Adsorption; Analysis; Analytical Chemistry; Biochemistry; Calibration; Cell culture; Cell number; Centrifugal filtration; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Drinking water; Drinking Water - microbiology; Elution; Enrichment; Epoxy Compounds - chemistry; Epoxy resins; Filtration; Food Science; Glycine; Health problems; Hydrogen-Ion Concentration; Hydrophobic and Hydrophilic Interactions; Hydrophobicity; Ionic interactions; Laboratory Medicine; Methods; Microscopy, Electron, Scanning - methods; Monitoring/Environmental Analysis; pH effects; Porosity; Pseudomonas aeruginosa; Pseudomonas aeruginosa - isolation & purification; Real-Time Polymerase Chain Reaction - methods; Research Paper; Ultrafiltration; Water analysis; Water purification; Water sampling
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-020-02956-3

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 10 3 . 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. Graphical abstract