title: Bacteriophage-Based Bioconjugates as a Flow Cytometry Probe for Fast Bacteria Detection.
abstract: Robust detection of bacteria can significantly reduce risks of nosocomial infections, which are a serious problem even in developed countries (4.1 million cases each year in Europe). Here we demonstrate utilization of novel multifunctional bioconjugates as specific probes for bacteria detection. Bifunctional magnetic-fluorescent microparticles are coupled with bacteriophages. The T4 bacteriophage, due to its natural affinity to bacterial receptors, namely, OmpC and LPS, enables specific and efficient detection of Escherichia coli bacteria. Prepared probes are cheap, accessible (even in nonbiological laboratories), as well as versatile and easily tunable for different bacteria species. The magnetic properties of the bioconjugates facilitate the separation of captured target bacteria from other components of complex samples and other bacteria strains. Fluorescence enables simple analysis. We chose flow cytometry as the detection method as it is fast and widely used for biotests. The capture efficiency of the prepared bioconjugates is close to 100% in the range of bacteria concentrations from tens to around 105CFU/mL. The limit of detection is restricted by flow cytometry capabilities and in our case was around 104CFU/mL.
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.
authors.forename: Marta
authors.initials: M
authors.lastname: Janczuk
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.
authors.forename: Łukasz
authors.initials: Ł
authors.lastname: Richter
authors.affiliation: Laboratory of Flow Cytometry, Medical Center of Postgraduate Education , Marymoncka 99/103, 01-813 Warsaw, Poland.
authors.forename: Grażyna
authors.initials: G
authors.lastname: Hoser
authors.affiliation: Department of Biomedical Systems and Technologies, Nalecz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences , Trojdena 4, 02-109 Warsaw, Poland.
authors.forename: Jerzy
authors.initials: J
authors.lastname: Kawiak
authors.affiliation: Department of Molecular Biology, University of Gdansk , Wita Stwosza 59, 80-308 Gdansk, Poland.|Phage Consultants , Partyzantów 10/18, 80-254 Gdansk, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Łoś
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Niedziółka-Jönsson
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.
authors.forename: Jan
authors.initials: J
authors.lastname: Paczesny
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences , Kasprzaka 44/52, 01-224 Warsaw, Poland.
authors.forename: Robert
authors.identifier: 0000-0002-3211-4286
authors.initials: R
authors.lastname: Hołyst
mesh_terms: D017122:Bacteriophage T4 / Q000378:metabolism*; D004926:Escherichia coli / Q000302:isolation & purification*; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D008863:Microspheres; D013997:Time Factors
chemical_list: D005456:Fluorescent Dyes