FIELD: biotechnology.
SUBSTANCE: technique for determining the film-forming function of pseudomonads based on mass spectrometry using the MALDI-ToF method. The studied pseudomonad cultures are sown in solid Mueller-Hinton agar medium, poured into Petri dishes using the exhaustion streak method in order to obtain single colonies. Next, the plating is incubated at a temperature from 36.5 to 37.5°C for 20 to 24 hours. Next, into the test tube with the HCCA matrix (α -Cyano-4-hydroxycirmamicacid) add 250 µl of OS standard solution: 50% acetonitrile, 47.5% water and 2.5% trifluoroacetic acid. The resulting suspension is stirred on a laboratory agitator at room temperature until the dry substance is completely dissolved. The biomaterial is then applied in a thin layer to a smooth target. Then cover with 1 μl of HCCA matrix solution and dry at room temperature. After complete drying, mass/charge peaks with values of 2348, 2361, 3171, 4174, 8889 Da are identified on a MALDI-ToF mass spectrometer.
EFFECT: expanding the functionality of the method, as well as reducing labor intensity by introducing the use of fewer reagents and reducing the time for preparing biological samples of bacteria, as well as increasing the accuracy of identification of microorganisms by detecting a pattern with an m/z value in the range of 2261, 2348, 2362, 3173, 4175, 8886 Da, responsible for biofilm formation in pseudomonads.
1 cl, 3 tbl, 1 ex
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Authors
Dates
2023-11-09—Published
2023-04-12—Filed