Basic Guidelines for Bacteriophage Isolation and Characterization

Theodor Bilharz Research Institute

Bibliographic Information

Authors: Samir S.

Journal: Recent Patents on Biotechnology

Publisher: Bentham Science Publishers

Publication Date: December 2023

Volume / Issue: Volume 17 / Issue 4

Pages: 312–331

ISSN: 18722083

DOI: 10.2174/1872208317666221017094715

Scopus: View on Scopus

PubMed: 36263478

Document Type: Review

Authors and Affiliations

Samir S., Department of Biochemistry and Molecular Biology, Theodor Bilharz Research Institute, Giza, Egypt

Abstract

The world is on the cusp of a post-antibiotic period. A century ago, before the advent of antibiotics, bacteriophage therapy was the treatment of choice for bacterial infections. Although bacteriophages have yet to be approved as a treatment in Western medicine, researchers and clinicians have begun to anticipate phage therapy. Bacteriophages are viruses that depend on bacterial cell metabolism to multiply. They offer a promising alternative to the use of antibiotics and an excellent antibacterial option for combating multi-drug resistance in bacteria. However, not every phage is suitable for phage therapy. In particular, prophages should not be used because they can lysogenize host cells instead of lysing them. To offer adequate therapeutic options for patients suffering from various infectious diseases, a wide selection of different phages is needed. While there is no evidence of direct toxicity induced by phage particles, it is crucial to study mammalian cell–phage interactions. This requires phage preparations to be free of bacterial cells, toxins and other compounds to avoid skewing host responses. Negative staining of purified viruses and electron microscopy remain the gold standard in the identification of bacteriophages. Interestingly, genomics has greatly changed our understanding of phage biology. Bacteriophage genome sequencing is essential to obtain a complete understanding of the bacteriophages' biology and to obtain confirmation of their lifestyle. Full genetic sequencing of bacteriophage will enable a better understanding of the phage-encoded proteins and biomolecules (especially phage lytic enzymes) involved in the process of bacterial cell lysis and death. Mass spectrometry can be used for the identification of phage structural proteins. The use of lytic phages as biocontrol agents requires the most appropriate and standard methods to ensure application safety. This review pursues recent research and methods in molecular biology for the isolation and characterization of phages to facilitate follow-up and implementation of work for other researchers. Patents related to this topic have been mentioned in the text. © 2023 Bentham Science Publishers.

Keywords

antibiotic alternatives; antibiotics; Bacterial infection; bacteriophage isolation; bacteriophage therapy; electron microscopy; genome sequencing; mass spectrometry; Animals; Anti-Bacterial Agents; Bacteria; Bacterial Infections; Bacteriophages; Humans; Mammals; Patents as Topic; Cytology; Electron microscopes; Genes; Metabolism; Molecular biology; Proteins; antibiotic agent; deoxycholic acid; enterotoxin; mitomycin; RNA 16S; antiinfective agent; Antibiotic alternative; Bacterial cells; Cell metabolism; Isolation and characterization; Phage therapy; Western medicines; antibiotic resistance; bacterial clearance; bacteriophage; biological control agent; CRISPR Cas system; cytolysis; cytotoxicity; double agar overlay; droplet digital polymerase chain reaction; drug delivery system; economic aspect; enzyme linked immunosorbent assay; Escherichia coli; flow cytometry; follow up; genomics; Haemophilus influenzae; high throughput sequencing; human; liquid chromatography-mass spectrometry; mass fragmentography; matrix-assisted laser desorption-ionization mass spectrometry; microbiological examination; multidrug resistance; nonhuman; patent; pH; phage display; polymerase chain reaction; practice guideline; protein interaction; Pseudomonas aeruginosa; Pseudomonas syringae; random amplified polymorphic DNA; reverse transcription polymerase chain reaction; Review; Streptococcus iniae; Streptococcus pneumoniae; transmission electron microscopy; Vibrio cholerae; virus cell interaction; virus characterization; virus isolation; virus purification; virus replication; whole genome sequencing; animal; bacterium; genetics; mammal; microbiology

Citation Information

Scopus Citations: 7

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