METHOD OF DETERMINING DEGREE OF EPIDEMIC HAZARD OF PATHOGENIC AND POTENTIALLY PATHOGENIC BACTERIA ISOLATED FROM WATER FOR VARIOUS USES Russian patent published in 2012 - IPC C12Q1/00 C12R1/385 C12R1/19 C12R1/22 C12R1/42 C12R1/445 C12R1/39 

FIELD: chemistry.

SUBSTANCE: adhesive activity index (A), invasive activity index (I) and toxic activity index (T) of pathogenic bacteria is determined on continuous cells of a tissue culture of buffalo green monkey (BGM) kidney cells. A dose containing 10⁷ colonies of bacterial cells is added to the culture monolayer of BGM cells and held at 37°C for 24 hours. The monolayer cells are washed 8 times with 10 ml of a physiological solution at pH 7.2. Index A is determined using a formula after first visually counting in Gorjaev's chamber living and dead cells stuck with bacterial growth and coloured by 0.1% trypan blue solution. Index (I) is determined using a formula after first replacing the culture medium in vials with a medium containing an antibiotic with concentration 50 mcg/ml and incubated in a temperature-controlled cabinet for 1 hour at 37°C. BGM cells are washed from bacteria 8 times with 10 ml of a physiological solution and lysed with 0.1% Triton X-100. Seeding of 0.1 ml is made from the obtained suspension based on the required cultivations per cup with Endo medium. Incubation is carried for 24 hours in a temperature-controlled cabinet at 37°C and the number of grown bacterial cells is counted. Index (T) is determined using a formula after first incubating bacteria in the BGM cell monolayer for 1 day. The suspension is centrifuged and filtered through membrane filters with pore diameter 0.22 mcm. The obtained substrate is put into a pure monolayer of continuous cells and the number of living and dead BGM cells is counted after one day.

EFFECT: invention enables to determine the degree of epidemic hazard of pathogenic bacteria isolated from water.

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