IDENTIFICATION METHOD OF BLUE PUS BACILLUS PSEUDOMONAS AERUGINOSA Russian patent published in 2015 - IPC G01N33/483 

FIELD: biotechnologies.

SUBSTANCE: identification method of blue pus bacillus Pseudomonas aeruginosa involves inoculation of the test material on a hard substrate, incubation of the inoculum under anaerobic conditions at the temperature of 37°C during 16-18 hours. The obtained bacterial mass in the amount of one bacteriological loop is placed in 300 mcl of a physiological solution and warmed-up at the temperature of 98-99°C during 20-30 minutes and centrifuged at 12000 revolutions per minute during 30 seconds. To a supernatant there added is colouring material for electrophoretic detection in the quantity of 0.5 mcl and 20 mcl is added to a well with a size of 4×1 mm 1.2% agarose gel on TAE-buffer with 10 mcl of 1% ethidium bromide. Besides, the amount of 3 mcl of the solution of standard DNA-marker 1 kb containing DNA fragments in the range of 250-10000 bp is added to the test well. Horizontal electrophoresis is performed during 15-20 minutes, and at detection on the obtained electrophoregramme of the test bacterial mass of three strips radiating in ultraviolet light, one of which corresponds to fragments of standard DNA-marker with the size of 10000 bp, the second one corresponds to fragments of standard DNA-marker with the size of 6000-8000 bp and the third strip at the end of the track in the form of a whisk, which corresponds to fragments of standard DNA-marker with the size of less than 750 bp, there identified is Pseudomonas aeruginosa in the test bacterial mass.

EFFECT: method allows quick and complete identification of pigment-shaping and non-pigment strains Pseudomonas aeruginosa in a test bacterial mass.

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