Method for detecting bacterial drug resistance in aquaculture

A detection method and aquaculture technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as time-consuming and labor-intensive, and achieve broad application prospects, strong practicability, high theoretical research significance and The effect of practical value

Inactive Publication Date: 2012-09-19
SUN YAT SEN UNIV +1
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AI Technical Summary

Problems solved by technology

Therefore, it is time-consuming and labor-intensive to evaluate resistant consumables by traditional microbial culture methods.

Method used

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  • Method for detecting bacterial drug resistance in aquaculture
  • Method for detecting bacterial drug resistance in aquaculture
  • Method for detecting bacterial drug resistance in aquaculture

Examples

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Embodiment 1

[0038] Example 1 Citrobacter freundii Citrobacter freundii drug allergy testing

[0039] Weigh chloramphenicol, trimethoprim, gentamycisulfate, florfenicol, sulfamethoxazole, sulfadiazine, tetracycline ( Tetracycline), Kanamycin (Kanamycin sulfate) 8 kinds of antibiotic drugs, add MHB medium to dilute the drugs, so that the final concentration of the drug solution is 1024 μg / ml, the drug solution is filtered through the filter membrane, and packed in sterile vials at -80°C save.

[0040] The isolated and purified Citrobacter freundii was inoculated in MHB medium and cultured overnight with shaking at 30°C. Adjust the concentration of the bacterial solution with MNB medium to make the final concentration 1×10 6 CFU / ml.

[0041] Add 200 μl of chloramphenicol solution with a final concentration of 1024 μg / ml to the 2nd well of the first row of a sterile 96-well microtiter plate; add 100 μl of MHB medium to the 3rd to 11th wells; use two-fold dilution method, use a 100μl p...

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Abstract

The invention discloses a method for detecting bacterial drug resistance in aquaculture, which comprises the following steps: (1) an antibacterial drug stock solution is prepared; (2) a bacterial liquid is prepared; (3) an antibiotic working solution is prepared from the antibacterial drug stock solution obtained in step (1); (4) a 96-well microtitration plate is placed in an incubator for incubation; and (5) data are read through a microplate reader for analysis. The method applies to bacterial drug resistance detections in aquaculture and other areas. In combination of the specification of the 96-well microtitration plate (8 multiply 12) and the drug concentration scope required by CLSI (Clinical and Laboratory Standards Institute), 8 categories of antibiotic drugs and 10 concentration gradients are selected for detection. When the method is in use, no flat plate is required, the steps of configuring a large amount of culture mediums, manufacturing drug sensitive paper scraps and measuring the inhibition zone are spared, and a small quantity of culture solutions is required. The method has the advantages of strong practicability, convenience, accuracy and stability in results, good repeatability and high sensitivity.

Description

technical field [0001] The invention relates to a high-throughput detection method for drug resistance of aquatic bacteria. It can be extended to the detection of drug resistance of other bacteria through different selection of resistant drugs. Background technique [0002] Antibiotics are medicines used to treat various bacterial infections or to suppress infection by pathogenic microorganisms. For a long time, antibiotics have played a vital role in human life and production, but repeated use of an antibiotic may cause pathogenic bacteria to develop drug resistance. [0003] my country is the world's largest aquaculture country, but in recent years, due to the intensification of aquaculture pollution, the quality of the aquaculture environment has continued to deteriorate, the variety of pathogenic organisms has increased and the speed of transmission has accelerated, and aquaculture diseases have become increasingly serious. The disease problems in aquaculture are especi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02
Inventor 黄志坚陈旭凌袁永青何建国
Owner SUN YAT SEN UNIV
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