Method for evaluating migration risk of antibiotic drug resistance gene

A technology for drug resistance genes and antibiotics, which is applied in biochemical equipment and methods, microbial determination/inspection, resistance to vector-borne diseases, etc. achieve high sensitivity

Active Publication Date: 2022-05-17
GUANGDONG UNIV OF TECH
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Problems solved by technology

[0003]Traditional research on horizontal transfer of drug-resistant genes is to select zygotes in liquid-solid medium containing antibiotic selection pressure by using filter membrane or liquid incubation method in vitro Dilution smear cou

Method used

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  • Method for evaluating migration risk of antibiotic drug resistance gene
  • Method for evaluating migration risk of antibiotic drug resistance gene
  • Method for evaluating migration risk of antibiotic drug resistance gene

Examples

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Example Embodiment

[0067] Embodiments

[0068] The present embodiment is a method for assessing the risk of antibiotic resistance gene migration, as follows.

[0069](1) Remove red fluorescent E. coli K12 and green fluorescent E. coli DH5α from the -80 °C refrigerator, activate and coat to check whether the fluorescent proteins of E. coli red and green fluorescent E. coli are normally expressed.

[0070] (2) The above two strains of E. coli were selected in the LB culture medium to be cultured to the logarithmic growth stage, of which the LB culture solution for the cultivation of E. coli green Escherichia coli DH5α contains 100 μg / mL ampicillin, 50 μg / mL kanamycin and 10 μg / mL tetracycline hydrochloride.

[0071] (3) Wash and dilute E. coli K12 and E. coli DH5α with normal saline or sterile phosphate buffer (0.1 mM) to 1010 CFU / mL of bacteria respectively to obtain pre-bonded bacterial solution.

[0072] (4) With E. coli red K12 as the recipient bacterium and E. coli green Escherichia coli DH5α as...

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Abstract

The invention discloses a method for evaluating the migration risk of antibiotic drug resistance genes. The method comprises the steps that a first target strain and a second target strain are provided, the first target strain has double fluorescence signals, and the second target strain has multi-antibiotic sensitivity; enabling the first target strain to be in contact with a second target strain in a sample to be detected under a condition suitable for jointing to obtain a zygote; and carrying out fluorescence detection on the zygotes. According to the method, the risk of antibiotic drug-resistant gene migration can be rapidly evaluated, the defect that time and labor are wasted in an existing migration frequency calculation method is effectively overcome, and meanwhile the risk of drug-resistant gene horizontal migration under different environmental conditions is rapidly evaluated. In addition, the double-fluorescence drug-resistant escherichia coli constructed by the invention is used as a donor bacterium for researching the horizontal conjugational transfer of the antibiotic drug-resistant gene, and the drug-resistant gene entering the intestinal tract of a plant or an aquatic organism can be quickly positioned, so that the drug-resistant gene transfer visualization is realized.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for assessing the migration risk of antibiotic resistance genes. Background technique [0002] With the widespread use of antibiotics, the number of antibiotic-resistant bacteria driven by antibiotic resistance genes (ARGs) is increasing, and other non-resistant bacteria can also acquire resistance through conjugation, transduction, and transformation. The water environment is usually a natural reservoir of drug-resistant bacteria and drug-resistant genes, and an important environmental medium for the preservation, amplification, transfer and diffusion of drug-resistant genes. A large number of reports have shown that hundreds of drug resistance genes or even multiple drug resistance genes have been detected in various surface waters, including hospital sewage, domestic sewage, sewage treatment plants, groundwater and even drinking water. [0003] The traditional horizontal...

Claims

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Application Information

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IPC IPC(8): G01N21/64C12Q1/10
CPCG01N21/6486C12Q1/10Y02A50/30
Inventor 许燕滨林晓君韩睿琪
Owner GUANGDONG UNIV OF TECH
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