METHOD OF MELIOIDOZE ANTIGEN ESTIMATE FOR PROTECTIVITY LEVEL Russian patent published in 2009 - IPC A61K39/02 G01N33/48 C12N5/00 

FIELD: chemistry.

SUBSTANCE: to estimate protectivity level of melioidoze antigens, they are used as antigens being cytokine inducers on peritoneal macrophages (PM) culture of mice BALB/c. In holes of a 96-holed pad, PMs 200 mcl (10⁵ cells) are introduced, then antigens are added to final concentration 10 mkg/ml, while saline is added in control holes. After incubation within 24 h at 37°C and 5% CO₂, supernatants are collected, sterilised through filters Millipore 0.22 mcm. Cytotoxic FNO-α activity in prepared monokines is evaluated on transplantable cell line L-929 culture distributed on 100 mcl (2·10⁵ cells) in holes of a 96-holed pad. After incubation within 24 h prepared monokines 100 mcl are added in three repeatabilities. To intensify FNO-α action, actinomycin D 10 mcl is added in the holes to final concentration 1 mkg/ml. After incubation within 24 h, cytotoxic FNO-α action is evaluated. FNO-α activity is expressed with a number of activity units reciprocal to monokine cultivation ensuring 50% destruction of L-929 cells in a hole ("ЦГЕ"50). FNO-α activity degree FNO-α in experiemnt and control shows protectivity level of melioidoze antigens.

EFFECT: method is characterised with responsiveness, simplicity, profitability, it is not associated with using living agents of special danger infections as an examination tool.

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