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Detection kit, detection method and application for Escherichia coli fluoroquinolone antibiotic resistance

An antibiotic resistance, fluoroquinolone technology, applied in the field of microbial resistance, can solve problems such as large errors and low measurement efficiency

Active Publication Date: 2020-12-01
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Aiming at the problems of low efficiency and large error of the existing drug susceptibility method, the present invention provides a detection kit for Escherichia coli fluoroquinolone antibiotic resistance

Method used

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  • Detection kit, detection method and application for Escherichia coli fluoroquinolone antibiotic resistance
  • Detection kit, detection method and application for Escherichia coli fluoroquinolone antibiotic resistance

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Embodiment 1

[0098] The present embodiment provides a kind of detection method that is used for Escherichia coli fluoroquinolone antibiotic resistance, and described detection method is high-throughput fluorescent quantitative PCR method, and described detection method comprises the following steps

[0099] Step 1, extracting a DNA sample from the sample to be tested;

[0100] After the Escherichia coli is isolated clinically from the colibacillosis cases, the DNA of the isolated pathogenic Escherichia coli is firstly extracted for further determination. SoilPure gene DNA Isolation Kit (Cat. No.: DN2701, Company: Aidlab, Country: China) was used for DNA extraction.

[0101] The steps are as follows: take 0.5mL bacterial liquid and centrifuge, discard the supernatant, add 0.5mL solution SUS, stir with a pipette tip and vortex briefly to help resuspend. Add 20 μL proteinase K (20 mg / mL) solution and briefly vortex to help mix. Add 120 μL of solution LYS, briefly vortex to mix, and incubate...

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Abstract

The invention relates to the technical field of drug resistance of microorganisms, and particularly discloses a detection kit and a detection method for drug resistance of Escherichia coli against fluoroquinolone antibiotics and an application thereof. In the invention, primers for sixteen genes of emr drug resistance pathway and mdt drug resistance pathway, related to fluoroquinolone, are designed; then with the 16S rDNA as reference genes, amplification is carried out with a high-throughput fluorescent quantitative PCR instrument; and through data processing, the differential expression multiple between the relative expression quantity of the fluoroquinolone related genes of the bacteria and the bacteria can be intuitively reflected. Compared with a drug sensitive test with rough determination of the drug resistance, the method is more scientific and precise.

Description

technical field [0001] The invention relates to the technical field of microbial drug resistance, in particular to a detection kit, detection method and application for Escherichia coli fluoroquinolone antibiotic resistance. Background technique [0002] The most commonly used method in clinical laboratory medicine is the K-B disc method. At present, the National committee forclinical laboratory standards (Nc CLS) Disc Diffusion Sensitive Experiment Subcommittee recommends the standard method based on the K-B disc method established by Bauer-Kirby. The method is based on the currently recognized standard experimental method. Paste the drug-sensitive paper (containing quantitative antibacterial drugs) on the specific part marked on the agar plate that has been inoculated with the bacteria to be tested. The principle is that the drug diffuses into the surrounding agar by its molecular diffusion force. As the distance from the drug source increases, The concentration of the dr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/6851C12Q1/10C12R1/19
CPCC12Q1/6851C12Q1/689C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 张铁王春光张石磊翟向和史万玉
Owner HEBEI AGRICULTURAL UNIV.