FIELD: microbiology.
SUBSTANCE: described is a method for determining pneumonia agents by the mass spectrometric method MALDI-TOF, which consists in the fact that a sputum sample is dissolved in ratio of 1:2 in isotonic sodium chloride solution, then stirred on a Vortex laboratory mixer for 2-3 minutes and centrifuged at 13000 rpm for 2 minutes. Obtained precipitate is mixed in an equal volume with 95 % ethanol, then mixed on a Vortex laboratory mixer for 2-3 minutes and centrifuged at 13000 rpm for 2 minutes. Obtained precipitate is stirred in ratio of 1:1 in mixture of 70 % formic acid and 50 % acetonitrile, then mixed on a laboratory Vortex mixer for 2-3 minutes and centrifuged at 13000 rpm for 2 minutes. Obtained precipitate is applied on a target, dried at temperature of 22±1 °C for 5-10 minutes, after which the points with the applied biomaterial are coated with 1 mcl of the HCCA matrix solution and left at temperature of 22±1 °C, after complete drying, identification is carried out on a MALDI-TOF mass spectrometer.
EFFECT: broader functional capabilities of the method, as well as reduced labour intensity due to the use of less reagents and reduced time for determining the pathogens of community-acquired pneumonia, as well as high accuracy of identifying microorganisms.
1 cl, 3 tbl, 2 ex
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