METHOD OF OBTAINING LYOPHILIZED PREPARATION FOR IMMUNOLOGICAL DIAGNOSTICS OF PREGNANCY AND INFERTILITY OF COWS AND HEIFERS Russian patent published in 2018 - IPC A61K35/16 

FIELD: veterinary science.

SUBSTANCE: invention relates to veterinary medicine, namely to methods for preparing preparations for the diagnosis of pregnancy and infertility of cows and heifers. For the preparation of a rabbit with a mass of 1.5–2 kg, 4-fold with an interval of 7 days, subcutaneously in the scapula area, is immunized with the antigen "Human chorionic gonadotropin", in a dose of 2,000 U, diluted in 1 ml of Freud's sterile adjuvant. 10 days after the last injection of the hormone, the serum is obtained by sedimentation, then 2 ml of the obtained immune serum diluted 1:5 in 1 % boric acid solution is added to 10 ml of a suspension of fine particles of 1–1.6 mcm alizarin. Mixture is kept at 37 °C for 2 hours and another 13 hours at a temperature of 4–8 °C, then the mixture is centrifuged to clarification, add to the precipitate 10 ml of a 2 % solution of normal rabbit serum in 0.5 % boric acid in saline solution and preserve with quinosol (merthiolate) in a ratio of 0.01 g of preserving agent per 100 ml of the liquid obtained after dissolving the precipitate. 6 % sucrose is added to the resulting mixture as the forming agent, cryoprotectant and stabilizer, then the resulting mixture is poured into sterile test tubes in a volume of 2 ml, placed in a freezer at a temperature of +20 °C. Temperature is then reduced to -20 °C and maintained for 1 hour, after that, the temperature is reduced from -20 °C to -30 °C with an exposure time of 1 hour, then the temperature is reduced from -30 °C to -40 °C with an exposure time of 1 hour, then the chamber is cooled to a temperature of -45…-50 °C, the holding time at this temperature is 3 hours, then for 3 hours the samples are kept in a freeze chamber at a temperature of -50 °C with the lowest pressure of 5.0×10^-2 mbar. After that, at a speed of 4 °C/h, the temperature is increased from -50 °C to -10 °C, further the temperature in the chamber is increased from -10 °C to +4 °C at speed of 2 °C/hour, and +4 °C to +20 °C at a speed of 5 °C/hour, then the preparation is dried for 3 hours at a temperature of +20 °C, then the test tubes are closed with sterile caps.

EFFECT: use of this method makes it possible to obtain a lyophilized preparation for the diagnosis of pregnancy.

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