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A New High-throughput Screening (HiTS) Method for Phages – Enabling Crude Isolation and Fast Identification of Diverse Phages with Therapeutic Potential

Nikoline S. Olsen, Niels Bohse Hendriksen, Lars H. Hansen, Witold Kot
doi: https://doi.org/10.1101/2020.03.27.011080
Nikoline S. Olsen
1Department of Environmental Science, Aarhus University, Frederiksborgvej 399, Roskilde, Denmark,
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  • For correspondence: niso@envs.au.dk
Niels Bohse Hendriksen
1Department of Environmental Science, Aarhus University, Frederiksborgvej 399, Roskilde, Denmark,
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  • For correspondence: niso@envs.au.dk
Lars H. Hansen
2Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark, ,
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  • For correspondence: lhha@plen.ku.dk wk@plen.ku.dk lhha@plen.ku.dk wk@plen.ku.dk
Witold Kot
2Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark, ,
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  • For correspondence: lhha@plen.ku.dk wk@plen.ku.dk lhha@plen.ku.dk wk@plen.ku.dk
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Abstract

Bacteriophage therapy and application of phages as biocontrol in plant production and food processing, all necessitates acquisition of suitable phages. Depending on purpose, the selection criteria of phage characteristics include lifestyle (lytic/lysogenic), host range, physical stability and absence of unwanted genetic traits such as integrases, antibiotic resistance or bacterial virulence factors. The exclusivity of antibiotic resistant clinical infections and possible development of phage-resistance instigates a need to continually build sizeable phage libraries and also be able to rapidly isolate and characterise novel phages of specified bacterial hosts. Current methods for phage isolation are both laborious and time consuming, suitable only for the isolation of a limited number of phages. Thus, we developed the High-Throughput Screening (HITS) method for phages for fast isolation and identification of potentially hundreds of distinct phages against single hosts. This scalable method enables screening of hundreds of samples, in multiple simultaneous setups with varying parameters increasing the likelihood of isolating multiple distinct phages specific for the given conditions. The efficiency of the method is emphasised by our screening of 200 environmental samples, resulting in the identification of an abundance of unique phage species lytic to Escherichia coli, Salmonella Enterica, Enterococcus faecalis and Pseudomonas aeruginosa.

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Posted March 29, 2020.
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A New High-throughput Screening (HiTS) Method for Phages – Enabling Crude Isolation and Fast Identification of Diverse Phages with Therapeutic Potential
Nikoline S. Olsen, Niels Bohse Hendriksen, Lars H. Hansen, Witold Kot
bioRxiv 2020.03.27.011080; doi: https://doi.org/10.1101/2020.03.27.011080
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A New High-throughput Screening (HiTS) Method for Phages – Enabling Crude Isolation and Fast Identification of Diverse Phages with Therapeutic Potential
Nikoline S. Olsen, Niels Bohse Hendriksen, Lars H. Hansen, Witold Kot
bioRxiv 2020.03.27.011080; doi: https://doi.org/10.1101/2020.03.27.011080

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