METHOD FOR DIFFERENTIATION OF MALLEUS AND MELIOIDOSIS EXITANTS WITH PRECIPITATING ANTISERUM

摘要

FIELD: biotechnology.;SUBSTANCE: immunodiffusion reaction in gel is carried out with living cultures of analyzed strains B.mallei and B.pseudomallei and obtained rabbit antiserum is used as immune serum against exocellular antigens (ECA) of strain Burkholderia thialandensis 264. Analyzed strains B.mallei and B.pseudomallei are seeded on plate with solid mutrient medium (F-agar "Difco"), incubated for 18 h at 37°C. In agar wells are punched nearly of grown plaques and abovementioned rabbit antiserum is introduced. Plates are exposed for 18 h at 24°C and reaction results are analyzed. Presence of precipitation lines between plaques of seeded cultures and antiserum indicates seeding of bacterium B.pseudomallei. Absence of such lines indicates seeding of bacterium B.mallei. Antiserum is obtained by rabbit immunization with extracellular antigens of strain B.thialandensis 264 which represent filtrates of swab of bacterial mass grown on solid nutrient medium coated with cellophane. Filtrates are sterilized in two steps on membrane filters (pore size 0.45 mum and 0.22 mum). Extracellular filtrate components are precipitated with acetone, dried and used for rabbit immunization. Serum activity is evaluated by immunodiffusion reaction in gel. Blood is sampled at serum titer of 1:32 or more.;EFFECT: simplified method of improved accuracy.;2 cl, 4 ex