Bacteriophage-based bioconjugates as a flow cytometry probe for fast bacteria detection

Marta Janczuk , Łukasz Richter , Grażyna Hoser , Jerzy Kawiak , Marcin Łoś , Joanna Niedziółka-Jönsson , Jan Paczesny , Robert Hołyst

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 105 CFU/mL. The limit of detection is restricted by flow cytometry capabilities and in our case was around 10(4) CFU/mL.
Author Marta Janczuk
Marta Janczuk,,
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, Łukasz Richter
Łukasz Richter,,
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, Grażyna Hoser
Grażyna Hoser,,
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, Jerzy Kawiak
Jerzy Kawiak,,
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, Marcin Łoś (FB / DMGB)
Marcin Łoś,,
- Department of Molecular Genetics of Bacteria
, Joanna Niedziółka-Jönsson
Joanna Niedziółka-Jönsson,,
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, Jan Paczesny
Jan Paczesny,,
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, Robert Hołyst
Robert Hołyst,,
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Journal seriesBioconjugate Chemistry, ISSN 1043-1802, (A 35 pkt)
Issue year2017
Vol28
No2
Pages419-425
Publication size in sheets0.5
DOIDOI:10.1021/acs.bioconjchem.6b00596
URL http://pubs.acs.org/doi/pdf/10.1021/acs.bioconjchem.6b00596
Languageen angielski
Score (nominal)40
ScoreMinisterial score = 35.0, 20-12-2017, ArticleFromJournal
Ministerial score (2013-2016) = 40.0, 20-12-2017, ArticleFromJournal
Publication indicators WoS Impact Factor: 2016 = 4.818 (2) - 2016=4.574 (5)
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
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