Title:Basic Guidelines for Bacteriophage Isolation and Characterization
Volume: 17
Issue: 4
Author(s): Safia Samir*
Affiliation:
- Department of Biochemistry and Molecular Biology, Theodor Bilharz Research Institute, Giza, Egypt
Keywords:
Bacterial infection, antibiotics, antibiotic alternatives, bacteriophage therapy, bacteriophage isolation, electron microscopy, genome sequencing, mass spectrometry.
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 multidrug
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.
