METHOD OF DETERMINING SENSITIVITY OF BACTERIA TO EFFECT OF LYSOZYME OR LYSOSTAPHIN Russian patent published in 2011 - IPC C12Q1/02 

FIELD: chemistry; biochemistry.

SUBSTANCE: invention relates to biotechnology and specifically to methods of determining degree of sensitivity of bacteria to damaging effect of bacteriolytic enzymes. The method involves growing bacteria on dense culture medium for 20-24 hours at temperature 37 °C. The obtained culture of bacteria is washed with 0.9% NaCl solution and centrifuged. After centrifuging, the bacteria cells are resuspended in a buffer whose pH is optimum for the enzyme with subsequent measurement of optical density of the suspension at wavelength 540 nm, setting the density on the level of 0.5 units and determination of the absolute quantity of bacteria in 1 ml of the suspension. 0.2 ml lysozyme with concentration of 10 mcg/ml or lysostaphin in concentration of 2.5 mcg/ml is added to 2.8 ml of the obtained suspension. The suspension of cells and enzyme solution are mixed. Optical density is measured every 10 second for 600 seconds from the moment the enzyme solution is added, with subsequent evaluation of bacteria sensitivity on the section with maximum rate of reaction for its characteristic with number of bacteria cells lysed in unit volume (ml) per unit time (sec).

EFFECT: invention enables to carry out sound correction of a system of treatments with a positive clinical result.

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