title: Application of immobilized synthetic anti-lipopolysaccharide peptides for the isolation and detection of bacteria.
abstract: The molecular detection of microorganisms in liquid samples generally requires their enrichment or isolation. The aim of our study was to evaluate the capture and pre-concentration of bacteria by immobilized particular cationic antimicrobial peptides, called synthetic anti-lipopolysaccharide peptides (SALP). For the proof-of-concept and screening of different SALP, the peptides were covalently immobilized on glass slides, and the binding of bacteria was confirmed by microscopic examination of the slides or their scanning, in case of fluorescent bacterial cells. The most efficient SALP was further tethered to magnetic beads. SALP beads were used for the magnetic capture of Escherichia coli in liquid samples. The efficiency of this strategy was evaluated using polymerase chain reaction (PCR). Covalently immobilized SALP were capable of capturing bacteria in liquid samples. However, PCR was hampered by the unspecific binding of DNA to the positively charged peptide. We developed a method for DNA recovery by the enzymatic digestion of the peptide, which allowed for a successful PCR, though the method had its own adverse impact on the detection and, thus, did not allow for the reliable quantitative analysis of the pathogen enrichment. Immobilized SALP can be used as capture molecules for bacteria in liquid samples and can be recommended for the design of the assays or decontamination of the fluids. For the accurate subsequent detection of bacteria, DNA-independent methods should be used.
authors.affiliation: Nanotechnology Unit, Fraunhofer Institute for Cell Therapy and Immunology, Perlickstrasse 1, 04103, Leipzig, Germany, natalia.sandetskaya@izi.fraunhofer.de.
authors.forename: N
authors.initials: N
authors.lastname: Sandetskaya
authors.forename: B
authors.initials: B
authors.lastname: Engelmann
authors.forename: K
authors.initials: K
authors.lastname: Brandenburg
authors.forename: D
authors.initials: D
authors.lastname: Kuhlmeier
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000378:metabolism*; D001419:Bacteria / Q000145:classification* / Q000302:isolation & purification*; D001431:Bacteriological Techniques / Q000379:methods*; D004926:Escherichia coli; D006801:Humans; D008070:Lipopolysaccharides / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D023181:Antimicrobial Cationic Peptides; D008070:Lipopolysaccharides