METHOD FOR PRODUCING ANTIGEN FOR SEROLOGICAL DIAGNOSIS OF CANINE BABESIOSIS

摘要

According to the invention, the puppies of the dogs at the age of 4-5 months are infected with 0.5 mL of the blood from the dogs with the spontaneous babesiosis. The dogs are exsanguinated when the parasitemia attains its maximum (not less than 10%). Glucigir solution at a 1:4 ratio is used as anticoagulant and preserving agent for prolongation of the term of blood suitability for processing. The plasma is removed by centrifugation at 3000 rpm for 15 min. The red blood cells are washed in the physiologic saline solution at 6000 rpm for 15 min (2-3 times). The precipitate of the erythrocytes is lysed by saponin at a ratio of 1:10000, the lysate is washed with ten-fold volume of saline solution by centrifugation at 6000 rpm for 30 min two or three times until the clarification of the supernatant. The upper layer of the grey deposit is transferred to the flask. The precipitate is homogenized in the flask with the beads by hand shaking. Sodium azide is used as a preserving agent at a ratioof 1:10000. The antigen preparation is stored at the temperature +2-4 DEGREE C.