METHOD FOR PRODUCING PURIFIED SOMATIC ANTIGEN OF Dirofilaria immitis

摘要

FIELD: medicine.;SUBSTANCE: invention refers to medicine and veterinary science and concerns a method for producing the purified antigen of Dirofilaria immitis. The presented method involves mechanical homogenisation, centrifugation of a homogenate, collection of a supernatant to be used as an antigen; the homogenisation involves a 2-cm head end of a mature female of Dirofilaria immitis placed in an aqueous solution of saccharose 0.25 M in a ratio of 1:3, frozen at a temperature of -18°C, that is followed by mechanical homogenisation and protein extraction in the aqueous solution of saccharose 0.25 M at 4°C for 12 hours, wherein 3 cycles of five 30-second ultrasonic homogenisations of the supernatant is performed at 70 kHz every 30 seconds at 0°C; the supernatant prepared after ultrasonic homogenisation is dissolved in cooled acetone at a temperature of 0°C in a ratio of 1:20 with exposition for 1 hour at a temperature of 4°C.;EFFECT: presented invention enables producing the high-sensitivity and specificity antigen and can be used in diagnosis of dirofilariasis in humans and animals.;2 tbl, 1 ex