PCR Assay for Rapid Taxonomic Differentiation of Virulent Staphylococcus aureus and Klebsiella pneumoniae Bacteriophages

Phage therapy is now seen as a promising way to overcome the current global crisis in the spread of multidrug-resistant bacteria. However, phages are highly strain-specific, and in most cases one will have to isolate a new phage or search for a phage suitable for a therapeutic application in existin...

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Bibliographic Details
Published in:International journal of molecular sciences 2023-02, Vol.24 (5), p.4483
Main Authors: Kornienko, Maria, Bespiatykh, Dmitry, Malakhova, Maja, Gorodnichev, Roman, Kuptsov, Nikita, Shitikov, Egor
Format: Article
Language:English
Subjects:
Antibiotics
Antimicrobial agents
Bacteria
Bacteriophages - genetics
Classification
Deoxyribonucleic acid
DNA
Drug resistance
E coli
Genera
Genes
Genomes
Klebsiella
Klebsiella pneumoniae
Klebsiella pneumoniae - genetics
Laboratories
Lysates
Multidrug resistance
Myoviridae - genetics
Phages
Phylogenetics
Polymerase Chain Reaction
Proteins
Staphylococcus aureus
Staphylococcus aureus - genetics
Taxonomy
Virulence
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Summary:Phage therapy is now seen as a promising way to overcome the current global crisis in the spread of multidrug-resistant bacteria. However, phages are highly strain-specific, and in most cases one will have to isolate a new phage or search for a phage suitable for a therapeutic application in existing libraries. At an early stage of the isolation process, rapid screening techniques are needed to identify and type potential virulent phages. Here, we propose a simple PCR approach to differentiate between two families of virulent phages ( and ) and eleven genera of virulent phages ( , , , , , , , , , and ). This assay includes a thorough search of a dataset comprising ( = 269) and ( = 480) phage genomes available in the NCBI RefSeq/GenBank database for specific genes that are highly conserved at the taxonomic group level. The selected primers showed high sensitivity and specificity for both isolated DNA and crude phage lysates, which permits circumventing DNA purification protocols. Our approach can be extended and applied to any group of phages, given the large number of available genomes in the databases.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms24054483