METHOD FOR DETERMINING BLOOD COAGULATION PROTEIN IN ANIMALS Russian patent published in 2019 - IPC G01N33/68 G01N33/543 

FIELD: medicine.

SUBSTANCE: invention relates to medicine and represents a method for determining von Willebrand factor, including using immunohistochemistry, characterized by that peripheral blood smears from animals fixed with 96 % alcohol, after 5-minute washing in distilled water is transferred into 3 % aqueous solution of hydrogen peroxide for 10 minutes at room temperature, then washed with distilled water and placed in 0.01 M phosphate-salt buffer with pH 7.4 for 5–10 minutes, 5 % bovine serum albumen is applied for 10 minutes, on one of the areas of the smear is applied primary polyclonal rabbit antibodies to VWF, and the second region of the smear is coated with a phosphate-salt buffer, placing the slides with smears into a moist chamber and placing in temperature 27 °C for 24 hours, then antibodies are washed off, the glasses are placed in a vessel with a fresh portion of buffer for 5–10 minutes, secondary rabbit antibodies are applied, put in a thermostat with temperature of 27 °C for 35 minutes, washing off the antibodies and applying the working solution of 3,3-diaminobenzidine, for 1–3 minutes the process is controlled to stop the reaction until a nonspecific background appears, then the preparations are washed in 2-3 portions of distilled water for 3–5 minutes in each, the dashes are stained with azure-eosin according to May-Grunwald or Romanovsky, after dehydration in the alcohol of the ascending fortress, the preparations are clarified in xylene and enclosed in polysterol, von Willebrand factor is determined by its precipitates of black-brown color in the form of fine granularity and numerous agglutinated thrombocytes.

EFFECT: invention provides quick and simple determination of von Willebrand factor in blood.

1 cl, 3 dwg, 3 ex