Please use this identifier to cite or link to this item: https://doi.org/10.21256/zhaw-18117
Title: A major-capsid-protein-based multiplex PCR assay for rapid identification of selected virulent bacteriophage types
Authors : Born, Yannick
Knecht, Leandra E.
Eigenmann, Mirjam
Bolliger, Michel
Klumpp, Jochen
Fieseler, Lars
et. al : No
Published in : Archives of virology
Volume(Issue) : 164
Issue : 3
Pages : 819
Pages to: 830
Publisher / Ed. Institution : Springer
Issue Date: 2019
License (according to publishing contract) : CC BY 4.0: Attribution 4.0 International
Type of review: Peer review (publication)
Language : English
Subjects : Bacteriophage; Capsid protein; Erwinia amylovora; Escherichia coli; Multiplex polymerase chain reaction; Phylogeny; Salmonella enterica; Virulence
Subject (DDC) : 570: Biology
Abstract: Bacteriophages represent a promising alternative for controlling pathogenic bacteria. They are ubiquitous in the environment, and their isolation is usually simple and fast. However, not every phage is suitable for biocontrol applications. It must be virulent (i.e., strictly lytic), non-transducing, and safe. We have developed a method for identifying selected types of virulent phages at an early stage of the isolation process to simplify the search for suitable candidates. Using the major capsid protein (MCP) as a phylogenetic marker, we designed degenerate primers for the identification of Felix O1-, GJ1-, N4-, SP6-, T4-, T7-, and Vi1-like phages in multiplex PCR setups with single phage plaques as templates. Performance of the MCP PCR assay was evaluated with a set of 26 well-characterized phages. Neither false-positive nor false-negative results were obtained. In addition, 154 phages from enrichment cultures from various environmental samples were subjected to MCP PCR analysis. Eight of them, specific for Salmonella enterica, Escherichia coli, or Erwinia amylovora, belonged to one of the selected phage types. Their PCR-based identification was successfully confirmed by pulsed-field gel electrophoresis of the phage genomes, electron microscopy, and sequencing of the amplified mcp gene fragment. The MCP PCR assay was shown to be a simple method for preliminary assignment of new phages to a certain group and thus to identify candidates for biocontrol immediately after their isolation. Given that sufficient sequence data are available, this method can be extended to any phage group of interest.
Departement: Life Sciences and Facility Management
Organisational Unit: Institute of Food and Beverage Innovation (ILGI)
Publication type: Article in scientific journal
DOI : 10.1007/s00705-019-04148-6
10.21256/zhaw-18117
ISSN: 0304-8608
1432-8798
URI: https://digitalcollection.zhaw.ch/handle/11475/18117
Published as part of the ZHAW project : Mechanisms of phage resistance in Erwinia amylovora
Appears in Collections:Publikationen Life Sciences und Facility Management

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