Method for Improving the Efficiency of Aminoglycoside Antibiotics in Killing Persisting Bacteria by Low Ion Shock

An aminoglycoside and antibiotic technology, applied in the fields of botanical equipment and methods, chemicals for biological control, biocides, etc., can solve the problems of bacterial resistance, long contact time, etc., to eliminate bacterial persistence , reduce side effects and improve efficiency

Active Publication Date: 2020-09-01
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Treating bacterial infections with antibiotics is a typical clinical and aquaculture treatment, but with the widespread use of antibiotics, some bacteria have developed drug resistance
In addition, in some chronic persistent infections and biofilm infections, the bacteria did not undergo drug resistance mutations, but required longer contact time when using antibiotic treatment, this phenomenon is related to the persistence of bacteria (persistence)

Method used

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  • Method for Improving the Efficiency of Aminoglycoside Antibiotics in Killing Persisting Bacteria by Low Ion Shock
  • Method for Improving the Efficiency of Aminoglycoside Antibiotics in Killing Persisting Bacteria by Low Ion Shock
  • Method for Improving the Efficiency of Aminoglycoside Antibiotics in Killing Persisting Bacteria by Low Ion Shock

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Low ion shock enhances the efficiency of aminoglycoside antibiotics in killing type Ⅰ Escherichia coli

[0025] 1. Activate the Escherichia coli standard strain (E.coli BW25113). Draw 1 μl of bacterial solution stored in a -80°C refrigerator with 40% glycerol, and inoculate it in 1ml M9 plus glucose liquid medium (configuration components: M9 medium, 5g / L glucose, 1.5g / L (NH 4 ) 2 SO 4 , 1mg / L VB 1 , PH=7.3), cultured on a shaker (220rpm) at 37°C to the plateau phase, re-diluted 1000 times and inoculated in 10ml M9 plus glucose liquid medium, and cultivated to the logarithmic phase (OD 600 = 0.5-0.6). Take two 5ml EP tubes, put 4ml bacterial solution in each tube and centrifuge (5000g, 4℃, 5min), remove the supernatant; resuspend and centrifuge with the same volume of 4℃ pre-cooled M9 liquid medium (5000g, 4℃, 5min ), remove the supernatant; use 1ml normal temperature M9 liquid medium (PH=7.3) to resuspend and centrifuge (5000g, 4°C, 5min), remove the supernatant; ...

Embodiment 2

[0037] Low ion shock enhances the efficiency of aminoglycoside antibiotics in killing type Ⅱ Escherichia coli

[0038] 1. Activate the Escherichia coli standard strain (E.coli BW25113). Take 1 μl of bacterial solution stored in a -80°C refrigerator with 40% glycerol, inoculate it in 1ml of MHB liquid medium (Mueller-Hinton broth), and culture it on a shaker (220rpm) at 35°C for 24 hours (plateau period 10 9 CFU / ml), re-diluted 1000 times and inoculated in 2ml MHB liquid medium, cultured on a shaker (220rpm) at 35°C for 24h (plateau stage 10 9 CFU / ml). Take a 1.5ml EP tube, absorb 1ml of bacterial liquid and centrifuge (13000g, 2min), remove the supernatant; resuspend with 1ml of M9 carbon-free liquid medium and transfer 9ml of M9 carbon-free liquid medium into sterilized shake flasks (i.e.: the bacterial solution is 10 8 Concentration of CFU / ml (Continue culturing in a medium using M9 carbon-free liquid medium as a substrate), treat on a shaker (220rpm) at 37°C for 5h and t...

Embodiment 3

[0050] Hypoionic shock enhances the efficiency of aminoglycoside antibiotics in killing type Ⅲ Staphylococcus aureus

[0051] 1. Activate the standard strain of Staphylococcus aureus (S.aureus ATC25923). Take 1 μl of bacterial solution stored in a -80°C refrigerator with 40% glycerol, inoculate it in 1ml of MHB liquid medium (Mueller-Hinton broth), and culture it on a shaker (220rpm) at 35°C for 24 hours (plateau period 10 9 CFU / ml), re-diluted 1000 times and inoculated in 2ml MHB liquid medium, cultured on a shaker (220rpm) at 35°C for 24h (plateau stage 10 9 CFU / ml). Take a 1.5ml EP tube, absorb 1ml of the bacterial liquid and centrifuge (13000g, 2min), remove the supernatant; use 1ml of YNB carbon source-free liquid medium (Yeast nitrogen base broth, without anamino acid) to resuspend and 9ml of YNB carbon source-free liquid The culture medium was transferred into sterilized shake flasks respectively (that is, the bacterial solution was divided into 10 8 The concentratio...

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Abstract

The invention discloses a method for improving the efficiency of killing retained bacteria by aminoglycoside antibiotics through low ion shock. The method comprises the following steps of: preparing antibiotics into an antibiotic pretreatment solution with a certain concentration by adopting an isotonic solution or a sterile aqueous solution, wherein the isotonic solution is a 0.3M glycerol solution or a 0.9% Nacl solution, and the sterile aqueous solution is double distilled water. The antibiotic pretreatment solution is used for carrying out resuspension treatment on bacterial liquid containing retained bacteria to be killed. The method can greatly improve the efficiency of killing the remained bacteria by the aminoglycoside antibiotics, effectively eliminate the retention of bacteria, reduce the tolerance of the bacteria, and reduce the dosage and the administration time on the premise of achieving the same treatment effect, thereby reducing the side effect of the antibiotics.

Description

technical field [0001] The invention belongs to the field of antibiotics and microorganisms, and in particular relates to a method for improving the efficiency of aminoglycoside antibiotics in killing persistent bacteria by low ion shock. Background technique [0002] Treating bacterial infections with antibiotics is a typical clinical and aquaculture treatment method, but with the widespread use of antibiotics, some bacteria have developed drug resistance. In addition, in some chronic persistent infections and biofilm infections, the bacteria did not undergo drug-resistant mutations, but required longer exposure time when using antibiotics, which is related to the persistence of bacteria. [0003] Persisters are a small portion of bacteria with phenotype drug resistance in the bacterial population. There is no drug resistance mutation in their own genes, but they are insensitive and resistant to antibiotics due to their low metabolism, no growth or slow growth. [0004] Nu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L2/02A61L2/18A01N43/16A01N47/44A01P1/00
CPCA01N43/16A01N47/44A61L2/02A61L2/18
Inventor 付新苗陈钟毓高媛媛
Owner FUJIAN NORMAL UNIV
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