Antibacterial and combined antibacterial activity of nitrosourea compound
A streptozotocin, positive bacteria technology, applied in the direction of antibacterial drugs, organic active ingredients, medical preparations containing active ingredients, etc.
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Embodiment 1
[0025] Embodiment 1 agar dilution method detects streptozotocin antibacterial MIC value
[0026] The tested strains were inoculated on agar culture dishes by drawing a line, and cultured overnight in a constant temperature incubator at 37°C. On the next day, pick at least 3 well-separated colonies with the same shape from the agar plate, inoculate them in appropriate broth medium, incubate at 35°C for 2-6 hours, and adjust the concentration of the bacteria solution to 0.5 McFarland turbidity using a turbidimeter. (1~2×10 8 CFU / mL) for use. Dilute the stock solution of streptozotocin with the corresponding diluent to various required concentrations in turn, add it to a 90mm sterile plate, then add the corresponding volume of MH agar with a graduated pipette, and fully mix the drug solution with the culture medium After homogenization, a drug-containing agar plate with a final concentration of streptozotocin in the range of 0.06-128 μg / mL was prepared for use. Dilute the 0.5 ...
Embodiment 2K-B
[0031] Embodiment 2K-B disc method combined drug susceptibility test
[0032] The tested strains were inoculated on agar culture dishes by drawing a line, and cultured overnight in a constant temperature incubator at 37°C. On the next day, pick at least 3 well-separated colonies with the same shape from the agar plate, inoculate them in appropriate broth medium, incubate at 35°C for 2-6 hours, and adjust the concentration of the bacteria solution to 0.5 McFarland turbidity using a turbidimeter. (1~2×10 8 CFU / mL), use a sterile cotton swab to dip in the bacterial solution, squeeze out the water lightly, spread it evenly on the MH agar medium, and dry the surface water for later use. The distance between the center points of the two discs is adjusted according to the drug sensitivity. If both drugs are sensitive, the distance is 24mm; if one of the two drugs is resistant and the other is sensitive, the distance is 15mm; if both drugs are resistant, the distance is 10mm. The MH ...
Embodiment 3
[0035] Embodiment 3 checkerboard method combined drug susceptibility test
[0036] The tested strains were inoculated on agar culture dishes by drawing a line, and cultured overnight in a constant temperature incubator at 37°C. On the next day, pick at least 3 well-separated colonies with the same shape from the agar plate, inoculate them in appropriate broth medium, incubate at 35°C for 2-6 hours, and adjust the concentration of the bacteria solution to 0.5 McFarland turbidity using a turbidimeter. (1~2×10 8 CFU / mL), and then diluted 20 times for use. The two compounds in the combined drug were respectively diluted horizontally (Drug A) and vertically (Drug B) in two round-bottom 96-well plates, and the diluted drug concentrations were 2 times the final experimental concentration, of which Add blank CAMH broth medium without drug after the last concentration gradient of each drug dilution. The diluted two drugs were completely combined at different concentrations, and adde...
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