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US 2011/0300528 A1Published application
Methods for Bacteriophage Design
OwnerSabah Abdel Amir Jassim, Ahmed Sahib Abdulamir +1inventors
InventorsSabah Abdel Amir Jassim, Ahmed Sahib Abdulamir, Fatima Abu Bakar
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Abstract
Methods for designing and breeding phages are described. The methods include methods to design phages for previously resistant bacterial strains. The methods described do not use genetic manipulation techniques.
Claims21
1-35. (canceled)
A method of modifying phage-host specificity, the method comprising incubating phages in a medium comprising of one or more of a chelating agent, detergent/surfactant, enzyme, lantibiotic, antibiotics and an agent which destroys cell walls.
A method of modifying phage-host specificity, the method comprising the steps of:—
- (a) obtaining large amounts of wild-type phages from at least one natural source by incubating the phages with bacterial hosts to obtain large numbers of phages,
- (b) removing bacterial host cells, to obtain a suspension of phages,
- (c) plating the suspension of phages from step (b) on a lawn of bacterial host cells,
- (d) assessing phage plaques to identify areas of highest phage activity,
- (e) isolating the areas of highest phage activity and isolating phages therefrom,
- (f) culturing the phages isolated in step (e) together with their host bacteria,
- (g) adding a viricidal mixture to the culture media of step (f) to remove free phages from the culture medium,
- (h) plating the viricidally-treated culture medium from step (g) onto a host bacterial lawn and identify plaques,
- (i) removing the plaques showing most virulent phage activity from the plate and isolate the phages therefrom,
- (j) incubating the phages obtained in step (i) in a medium comprising of one or more of a chelating agent, detergent/surfactant, enzyme, lantibiotic, antibiotics and an agent which destroys cell walls,
- (k) isolating the bacteriophages of step (j) and incubating them in a growth medium,
- (l) assessing the infectivity of the bacteriophages of step (k) and culturing those whose specificity has been modified,
- (m) storing the bacteriophages cultured in step (l).
10 more elements
A method of modifying phage-host specificity, the method comprising the steps of:—
- (a) obtaining large amounts of wild-type phages from at least one natural source by incubating the phages with bacterial hosts to obtain large numbers of phages,
- (b) removing bacterial host cells, to obtain a suspension of phages,
- (c) plating the suspension of phages from step (b) on a lawn of bacterial host cells,
- (d) assessing phage plaques to identify areas of highest phage activity,
- (e) isolating the areas of highest phage activity and isolating phages therefrom,
- (f) culturing the phages isolated in step (e) together with their host bacteria,
- (g) adding a viricidal mixture to the culture media of step (f) to remove free phages from the culture medium,
- (h) plating the viricidally-treated culture medium from step (g) onto a host bacterial lawn and identify plaques,
- (i) removing the plaques showing most virulent phage activity from the plate and isolate the phages therefrom,
- (j) incubating the phages obtained in step (i) in a medium comprising of one or more of a chelating agent, detergent/surfactant, enzyme, lantibiotic, antibiotics and an agent which destroys cell walls,
- (k) isolating the bacteriophages of step (j) and incubating them in a growth medium,
- (l) assessing the infectivity of the bacteriophages of step (k) and culturing those whose specificity has been modified,
- (m) storing the bacteriophages cultured in step (l).
- wherein step (d) further comprises the steps of assessing the biokinetics of the phage by
- (i) taking a sample of phage from step (c) and adding it to a bacterial culture,
- (ii) incubating the phage and bacteria together,
- (iii) exposing the mixture of phage and bacteria to a viricidal agent, in the incubation vessel,
- (iv) adding a surfactant the mixture in the incubation vessel and further incubating it, and
- (v) adding culture broth to the incubation vessel and incubating prior to plating on a bacterial lawn and assessment of plaque morphology.
16 more elements
Dependencies are inferred from the claim text.