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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 counting, although the whole process is simple to operate, it takes a long time and cannot be visualized, so it is impossible to quickly analyze the migration risk of drug resistance genes under different water quality environmental conditions

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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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Embodiment

[0068] This example is a method for assessing the risk of antibiotic resistance gene transfer, as follows.

[0069] (1) Take out red fluorescent Escherichia coli K12 and green fluorescent Escherichia coli DH5α from the -80°C refrigerator, apply after activation, and check whether the fluorescent proteins of red fluorescent Escherichia coli and green fluorescent Escherichia coli are normally expressed.

[0070] (2) Select the above two strains of Escherichia coli and culture them to logarithmic growth phase in LB culture medium, in which the LB culture medium for cultivating Escherichia coli DH 5α contains 100 μg / mL ampicillin, 50 μg / mL kanamycin and 10 μg / mL tetracycline hydrochloride.

[0071] (3) Wash and dilute Escherichia coli K12 and Escherichia coli DH 5α with physiological saline or sterile phosphate buffer (0.1 mM) until the number of bacteria reaches 1010 CFU / mL respectively to obtain a preconjugated bacterial solution.

[0072] (4) Red Escherichia coli K12 was used ...

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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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