Fast drug sensitivity detection kit for aquatic product vibrio

A kit and drug-sensitivity technology, which is applied in the field of rapid drug-sensitivity detection kits for Vibrio aquaticus, can solve the problems of cumbersome operation, heavy workload, and long cycle, and achieve the effect of shortening time and simple detection operation

Active Publication Date: 2013-04-03
MARINE BIOLOGY INST OF SHANDONG PROVINCE
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AI Technical Summary

Problems solved by technology

[0003] At present, in the field of aquatic bacterial disease prevention and control, the drug susceptibility testing methods mainly use the K-B disk method and broth dilution method, of which the K-B disk method generally must be carried out under the conditions of a professional microbiological laboratory, and requires personnel with professional knowledge to operate Completed, there are problems such as cumbersome operation, heavy workload, long

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  • Fast drug sensitivity detection kit for aquatic product vibrio
  • Fast drug sensitivity detection kit for aquatic product vibrio
  • Fast drug sensitivity detection kit for aquatic product vibrio

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Preparation of Vibrio aquaticus drug susceptibility rapid detection plate

[0037] 1. Drug configuration

[0038]According to the types of commonly used aquatic drugs and the list of banned drugs, the types of tested drugs were determined to be ampicillin, florfenicol, amikacin, tetracycline, oxytetracycline, doxycycline, polymyxin B, cotrimoxazole, Norfloxacin, enrofloxacin, rifampicin, a total of 11 kinds. According to the characteristics of different drugs, different solvents are used to accurately prepare the concentrations of 5.12mg / ml, 3.2mg / ml, 10.24mg / ml, 1.6mg / ml, 1.6mg / ml, 1.6mg / ml, 2.56mg / ml, 6.4mg / ml, 6.4mg / ml, 6.4mg / ml, 6.4mg / ml stock solution of antibacterial drugs, including ampicillin, doxycycline hydrochloride, amikacin, oxytetracycline hydrochloride, tetracycline hydrochloride, polymyxa Element B was prepared with ultrapure water; florfenicol and rifampicin were dissolved in methanol and diluted with ultrapure water to prepare; norfloxacin...

Embodiment 2

[0050] Embodiment 2: the preferred of rapid enrichment liquid

[0051] In order to realize the rapid growth of Vibrio and shorten the process of drug susceptibility testing, on the basis of Mueller-Hinton (MH) medium formula, it was improved to make bacteria enrichment solution. The improved formula was: MH0 (beef extract powder 6 g, Soluble starch 1.5 g, casein hydrolyzate 17.5 g, NaCl 15 g, 2.8 mg CaCl 2 , 4 mg MgCl 2 , distilled water 1 L), MH1 (beef dipping powder 6 g, soluble starch 1.5 g, casein hydrolyzate 17.5 g, NaCl 15 g, 2.8 mg CaCl 2 , 4 mg MgCl 2 , 10% fish broth, distilled water 1 L) and MH2 (beef dipping powder 6 g, soluble starch 1.5 g, casein hydrolyzate 17.5 g, NaCl 15 g, 2.8 mg CaCl 2 , 4 mg MgCl 2 , fish peptone 10 g, distilled water 1 L). In order to test the multiplication effect of different enrichment solution formulas on Vibrio parahaemolyticus, Vibrio alginolyticus and Vibrio splendidius, respectively draw 1 ml of the overnight cultured three bac...

Embodiment 3

[0053] Example 3: Vibrio parahaemolyticus is tested for drug susceptibility using a rapid detection microplate for Vibrio parahaemolyticus

[0054] 1) Pick a single colony of Vibrio parahaemolyticus VP-3 (the pathogenic bacterium of turbot) from the solid medium plate, inoculate it in the enrichment medium MH2 medium, and place it in a 28°C incubator for shaking culture ( Rotational speed: 120rpm) until turbidity is visible to the naked eye.

[0055] 2) Adjust the concentration of the enrichment solution to 0.5 wheat degrees with the test culture solution, and then dilute it 1500 times to make the concentration of the bacteria reach 1×10 5 A / ml is used for drug susceptibility testing. The formula of the detection culture medium described therein is: beef extract powder 6g, soluble starch 1.5g, casein hydrolyzate 17.5g, NaCl 15g, 2.8 mg CaCl2, 4 mg MgCl2, distilled water 1L.

[0056] 3) Add 2 drops (100 μl) of detection culture solution to each microwell in the quality contro...

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Abstract

The invention relates to a fast drug sensitivity detection kit for vibrio, which can be applied to an aquiculture site. According to the invention, the detection kit comprises a detection plate, a thallus collection tool, bacterium enrichment liquid, a drug sensitivity detection culture medium, a turbidimetry tube, an inoculation burette and a colorimetric card, wherein the detection plate comprises inspection holes enveloping drugs and color indicators for drug sensitivity detection, and the color indicators are Alamar Blue. According to the invention, fish peptone, Ca2+ and Mg2+ are added based on the MH culture medium and the concentration of NaCl is adjusted, and the culture medium is used as the bacterium enrichment liquid of vibrio, which can effectively promote the growth of the vibrio and enables the bacterium enrichment liquid to reach the required concentration in a short time, so as to shorten the time of drug sensitivity detection. At the same time, a blue dye, namely the Alamar Blue, which is safe and nontoxic to the cells is used; the Alamar Blue has no toxic effect on cells, and can be enveloped in the micropores of the drug sensitivity detection plate directly without influencing the growth of the bacteria, so that the detection operation is simpler and more convenient and quick; and the proliferation dynamic of the bacteria can be continuously monitored.

Description

technical field [0001] The invention belongs to the technical field of in vitro diagnosis of aquatic pathogenic microorganisms, and in particular relates to a rapid drug sensitivity detection kit for aquatic Vibrio. Background technique [0002] With the rapid development of aquaculture in recent years, the problem of aquaculture animal diseases has become increasingly prominent, among which bacterial diseases have caused huge losses to the fishery economy worldwide. Vibrio is a large group of Gram-negative bacteria with short, polar flagella, motile, non-spore-shaped, short rods bent into arcs, belonging to the family Vibrio and the genus Vibrio. Vibrio is widely distributed in nature, mostly in water bodies. A large number of studies have shown that a variety of Vibrio can cause epidemics and explosive deaths of commercial animals such as fish, shellfish, and crustaceans in seawater, mainly including Vibrio parahaemolyticus, Vibrio brilliant, Vibrio anguillarum, and Vibri...

Claims

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Application Information

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IPC IPC(8): C12M1/34C12Q1/20C12R1/63
Inventor 刁菁杨秀生叶海斌薛莲盖春蕾
Owner MARINE BIOLOGY INST OF SHANDONG PROVINCE
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