Micromolecular substance for improving sensitivity of bacteria to antibiotics
An antibiotic, susceptibility technology, used in the field of medicine
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Embodiment 1
[0035] Screening of Antibiotic-resistant Escherichia coli K12 BW25113
[0036] Escherichia coli ( Escherichia coli , E. coli )) is the most common type of bacteria in the intestinal tract of humans and animals, mainly parasitic in the large intestine, accounting for about 1% of intestinal bacteria. Detection of Escherichia coli starter bacteria by two-fold dilution method ( Escherichia coli The minimum inhibitory concentration (MIC) of K12 BW25113) against 10 antibiotics. will be 10 5 The colony-forming unit / mL starter bacteria were cultured in LB liquid medium containing 1 / 2 minimum inhibitory concentration of antibiotics at 37°C and passed down for 10 times, and the minimum inhibitory concentration of the obtained monoclonal was determined. It was found that the minimum inhibitory concentrations of the selected 10 bacteria to the 10 antibiotics were 64 times the minimum inhibitory concentrations of the respective starting bacteria (see Table 1), indicating that 10 ...
Embodiment 2
[0041] Glutamine is a key biological target of ampicillin-resistant Escherichia coli
[0042] 1. Preparation of GC-MS samples of ampicillin-resistant Escherichia coli
[0043] 1) Preparation of bacterial samples: Pick the Escherichia coli starter bacteria from the solid LB plate ( Escherichia coli K12 BW25113) and a single colony of ampicillin-resistant bacteria (see Example 1 for the specific method of cultivating drug-resistant bacteria), inoculate in 5 ml of LB medium, culture at 200 rpm at 37°C for 16 hours; press 1:100 (v / v) Inoculate in 100 ml of LB liquid medium and culture at 37°C until the OD600 value is 1.0.
[0044] 2) Preparation of GC-MS samples: take 10 ml of bacterial liquid, quickly add 5 ml of frozen methanol to quench the sample, so as to stop the metabolic process of the cells. The bacterial cells were collected by centrifugation at 8000 rpm at 4°C for 3 minutes to remove as much water as possible, and the bacterial cells were stored in 2 ml of cold ...
Embodiment 3
[0052] Glutamine downregulation is ubiquitous among Escherichia coli antibiotic-resistant strains
[0053] 1. Preparation of GC-MS samples of Escherichia coli resistant to various antibiotics
[0054] The starter bacteria of Escherichia coli ( Escherichia coli K12 BW25113) and its 10 resistant E. coli strains: GEN-R, AMP-R, BLFX-R, KAN-R, NA-R, CAZ-R, ER-R, CAP-R, SM-R and TET -R, see example 2.1 for sample preparation process
[0055] 2. Down-regulation of glutamine is ubiquitous in Escherichia coli resistant to various antibiotics
[0056] The prepared samples of 10 drug-resistant bacteria were loaded on GC-MS to analyze the metabolites. According to the retention time of glutamine in the GC-MS total ion chromatogram, the peak area data of each drug-resistant bacteria glutamine peak and the internal standard peak were obtained, and the relative peak area (ratio to the internal standard peak) was used to represent the metabolites content. The content of glutamine...
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