METHOD OF EXTRACTING VIRULENT BACTERIOPHAGES STAPHYLOCOCCUS AUREUS Russian patent published in 2020 - IPC C12N7/00 A61K35/76 

FIELD: medicine; biology; biotechnology.

SUBSTANCE: method for isolating virulent bacteriophages involves the following steps: a biofilm-forming Staphylococcus aureus isolate recovered from a removed tonsil of a patient with chronic adenoiditis or chronic tonsillitis is inoculated on the surface of sterile LB agar, incubated at 36–37 °C for 20–24 hours, grown colonies are selected, suspended in sterile LB-broth, suspension is incubated at 36–37 °C for 2–2.5 hours, then mitomycin C is added in amount of 1–10 mcg to the pre-incubated suspension, then additionally incubated with periodic shaking, then added to suspension chloroform taken in amount of 0.5–0.6 ml, followed by successively incubating obtained suspension at room temperature for 20–30 minutes, centrifuging the incubated suspension at 4,000–5,000 rpm for 20–30 minutes, supernatant is taken into a sterile test tube, incubated at room temperature for 1–2 hours, then sown according to Grazia in dilutions of 10^-2-10^-8 per daily culture of strain Staphylococcus aureus, free from induced bacteriophages, inoculation is successively incubated at 36–37 °C for 18–24 hours, selecting isolated plaques, suspending them in a sterile physiological solution, then added to the suspension chloroform taken in amount of 0.5–0.6 ml, the obtained suspension is again incubated, centrifuged and the supernatant is taken off.

EFFECT: method enables higher efficiency of extraction of natural virulent bacteriophages Staphylococcus aureus active against staphylococcus biofilms.

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