BACTERIOPHAGES PRODUCTION METHOD Russian patent published in 2017 - IPC C12N7/00 A61K35/76 G01N21/64 

FIELD: biotechnology.

SUBSTANCE: method comprises culturing bacterial cells of host strain in the absence of extraneous microflora, phage lysate preparation and purification by precipitation and/or filtration. After 30-120 minutes after culturing initiation, at the optimum growth temperature for the culture of rapidly growing host strain, every 30-60 minutes, smears are made from the host strain culture from the surface of solid nutrient medium or from a liquid culture medium. The smears are stained with a solution of acridine orange in the final concentration of 0.001% to 0.02% or acridine yellow solution to the final concentration of 0.01% to 0.2%. The stained smear is microscoped in fluorescence microscope. Time is set for inoculating the mother bacteriophage at achievement in the stained smear of not less than 50% in relation to the total host strain cells proportion of orange acridine stained cells, fluorescing with orange or red shades, or the proportion of yellow acridine stained cells fluorescing with yellow or orange shades, at achevement of less than 10% in relation to the total host strain cells proportion of cells with non-uniform fluorescence that is paired pole adjoining cells in the form of rods with irregular or spherical cytoplasm fluorescent and spherical or ellipsoidal cells with slightly fluorescenting central section. Then mother bacteriophage is inoculated.

EFFECT: stability of achieving high bacteriophage titer upon phage lysate production in a changing nutritive medium formulations, and increase in the range of variations of the indicator values of the host strain and bacteriophage culturing.

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