High-throughput rapid antimicrobial susceptibility test kit for veterinary clinical application

A detection kit and drug susceptibility technology, applied in the field of veterinary detection reagents, can solve problems such as time-consuming, inability to test antibiotic susceptibility, complicated operation steps, etc., and achieve the effects of intuitive results, easy observation, enrichment, and easy operation.

Pending Publication Date: 2018-11-23
苏州艾可瑞动物检测技术服务有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have some disadvantages, such as cumbersome operation steps, long time-consuming and inability to conduct drug susceptibility tests on a large number of antibiotics at one time.

Method used

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  • High-throughput rapid antimicrobial susceptibility test kit for veterinary clinical application
  • High-throughput rapid antimicrobial susceptibility test kit for veterinary clinical application
  • High-throughput rapid antimicrobial susceptibility test kit for veterinary clinical application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0144] Based on the sterilized 1640 medium, LB and TSB medium respectively, add 1% sterile calf serum and 1‰ sterile NAD. Inoculate clinically common Escherichia coli, Haemophilus parasuis (HPS) and Actinobacillus (APP) as the research objects, measure the OD600 absorbance value every 1 hour, measure each test tube three times, and draw the growth curve by taking the average value , to select the medium suitable for the rapid growth of most bacteria. Such as figure 1 As shown, the growth rate of Haemophilus parasuis on the 1640-based rapid enrichment medium is faster, it is easier to observe the enrichment, and at the same time, the influence of the color of LB and TSB medium on the results is reduced. Actinobacillus pleuropneumoniae and Haemophilus parasuis had the same trend. For E. coli, there was no difference between the 1640-based rapid enrichment medium and the LB medium.

Embodiment 2

[0146] Taking the Cmax of the drug in the blood after intramuscular injection of pigs at the normal dosage, the Cmax of the drug was coated on a 96-well ELISA plate with 2 times the Cmax and 1 times the Cmax respectively, and then freeze-dried to prepare a drug sensitivity detection plate.

[0147] Taking Escherichia coli, Actinobacillus pleuropneumoniae and Haemophilus parasuis as objects, the kit of the invention is used to detect the antibiotic sensitivity of Escherichia coli, Actinobacillus pleuropneumoniae and Haemophilus parasuis. Escherichia coli, Actinobacillus pleuropneumoniae and Haemophilus parasuis were respectively cultured on 1640-based rapid growth medium for 1 hour, 3 hours and 4 hours, and then 100 μL (2 drops) was added to the drug-sensitive plate. After incubating at 37° C. for 16 hours, 50 μL (1 drop) of 0.2% resazurin sterile solution was added to each well, and the results were read after incubating at 37° C. for 1.5 hours. The susceptibility results of v...

Embodiment 3

[0149] a. Use sterile cotton swabs to collect disease materials (in order to shorten the culture time of enrichment, collect as much as possible), add to the enrichment medium, and incubate at 37°C for 2-6h, when the colorless medium becomes slightly turbid ( About 0.5 McFarland turbidity, Figure 5 ), proceed to the next step; for stool samples, let stand for 30 minutes after collection, take 50-100 μL (1-2 drops) of the supernatant and add it to the enrichment medium; for blood samples, after standing for 30 minutes, take 50 μL of serum -100μL (1-2 drops) was added to the enrichment medium.

[0150] b. Add 100 μL (2 drops) of the cultured bacterial solution to each well of the coated drug-sensitivity plate, and incubate at 37°C for 16 hours;

[0151] c. Take out the drug sensitivity plate, add 50 μL (1 drop) of 0.2% resazurin sterile solution to each well, incubate at 37°C for 1.5 hours and read the results;

[0152] d. Judgment of drug susceptibility results: blue means c...

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Abstract

The invention discloses a high-throughput rapid antimicrobial susceptibility test kit for veterinary clinical application and a method. The kit comprises an independently-packaged enrichment medium, an antimicrobial susceptibility test plate and reaction liquid; the enrichment medium is selected from a sterile 1640 medium or an MH medium with 1% sterile neonatal bovine serum and 0.1% sterile NAD added; the antimicrobial susceptibility test plate is a reaction plate coated with a plurality of antibiotics, each hole of the reaction plate is coated with an antibiotic, and the concentration of theantibiotics is 1-2 times of the maximum plasma concentration after the clinical administration of the antibiotics; and the reaction liquid is a 0.2% resazurin sterile solution. According to the kit and the method, an antimicrobial susceptibility test is carried out, the time for the result is short, more drugs are tested at a time, the kit and the method are easy to operate, and special instruments and equipment are not needed; the test process is a liquid-phase reaction, more accords with the physiological environment and is close to the actual production; and the result is visual, specifically, blue represents susceptible, red represents susceptible, and the closer the color is to blue, the more susceptible a drug is.

Description

technical field [0001] The invention belongs to the field of veterinary detection reagents and relates to an antibiotic susceptibility detection kit for veterinary clinical use. Background technique [0002] Antibacterial drugs have played a very important role in the control of bacterial infectious diseases, but due to the unscientific and blind abuse of antibacterial drugs in the breeding process, many pathogenic bacteria have developed drug resistance, making antibacterial drugs less effective for bacterial diseases. The control effect is getting worse and worse, which not only causes the waste of medicine, but also delays the disease, causing great economic losses to the farmers. [0003] In vitro antimicrobial susceptibility test, referred to as drug susceptibility test (AST), refers to the test for measuring the antibacterial or bactericidal ability of drugs in vitro. In vitro drug susceptibility test is an indispensable test method for rational and effective use of a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/18
CPCC12Q1/18
Inventor 何成华樊彦红章雪明胡仁莉
Owner 苏州艾可瑞动物检测技术服务有限公司
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