FIELD: biotechnology.
SUBSTANCE: disclosed is a method for obtaining a brucellosis diagnosticum, which involves washing a two-day culture of brucella strain Brucella abortus 19 BA with 0.9% sodium chloride, bringing to concentration of 20-30 billion m.c. in 1 ml of suspension, inactivation of microbial suspension with 1% formalin solution at 22 °C for 2 hours, followed by washing and centrifugation, adding 0.9% sodium chloride solution, 0.04 % Carazzi hematoxylin solution, stirring, heating for 16-18 hours to 56 °C, centrifugation, separation of supernatant liquid, resuspension of obtained sediment in 0.9% solution of sodium chloride, centrifugation, addition to sediment of 0.1% solution of formalin and stabilizer in amount of 0.1 ml per 1 ml of obtained diagnosticum, selected from the group: 1.5% solution of polyvinylpyrrolidone and 7% solution of sucrose in ratio of 1:1 or 0.03% of carbonate buffer and 3% solution of sucrose in ratio of 1:1 and lyophilization, including stages of freezing and drying, wherein freezing of diagnosticum is carried out on pre-cooled to minus 45±5 °C shelves of lyophilic unit, freezing rate 8-10 °C/min, freezing time 3±0.25 h, and immediately after the end of freezing, the heating mode is set so that the temperature of the diagnosticum passes zero mark 0 °C to 10th hour from the beginning of drying, with the final temperature of additional drying of diagnosticum 30±2 °C, vacuum during drying at the stage of removing unbound water from 26 Pa to 10 Pa, and during additional drying 5±3 Pa.
EFFECT; invention provides an increase in the sensitivity of the diagnosticum and an increase in its shelf life.
1 cl, 2 ex
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Authors
Dates
2024-12-11—Published
2023-12-18—Filed