METHOD OF DIFFERENTIATION OF PLAGUE AND PSEUDOTUBERCULOSIS PATHOGENS AS PER N-ACETYL-BETA-D-GLUCOSAMINIDASE ACTIVITY Russian patent published in 2015 - IPC C12N1/20 C12Q1/04 G01N21/64 

FIELD: bioengineering.

SUBSTANCE: method of differentiation of plague and pseudotuberculosis pathogens as per N-acetyl-beta-D-glucosaminidase activity provides for production of the suspension of agarinic culture of studied bacteria in concentration (1-5)×109 microcolonies, preparation of synthetic substrate, for this 4-methylumbelliferil-N-acetyl-β-D-glucosaminide is used in amount of 50 mcM. The substrate is dissolved in 2 ml of dimethylformamide. From the produced solution 0.6 ml are taken, and 9.4 ml of 0.1 M phosphate buffer pH 7.4 is added. 20 mcl of prepared solution are mixed with droplet 0.05 ml of suspension of agarinic culture of bacteria in the physical solution in Petri dish. The obtained mixture is incubated for 10-20 minutes at 37°C, then reaction is terminated by addition of 5 mcl of 10 n. alkali solution. Differentiation in UV rays of transilluminator is performed at 366 nm. Bright fluorescence of blue colour is evidence of positive result of the reaction, and confirms that studied strain hydrolyses the prepared substrate and belongs to Yersinia pseudotuberculosis. Absence of incandescence confirms the strain belonging to Yersinia pestis.

EFFECT: invention ensures express diagnostics and differentiation of said bacteria, namely for 10-20 minutes.

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