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A method to identify whether Vibrio harveii is pathogenic

A technology of vibrio harveyi and pathogenicity, which is applied in the field of identifying whether vibrio harveyi is pathogenic, and can solve the problem that there is no report on the detection method of the strength of vibrio harveyi's pathogenicity, and achieves Avoid individual sensitivity differences, shorten the experimental period, and facilitate the detection of pathogenicity

Active Publication Date: 2020-09-01
NINGBO UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the reported detection methods of Vibrio harveyi, all focus on the qualitative aspect of Vibrio harveyi, but there is no report on the pathogenicity of Vibrio harveyi and the detection method of its pathogenic ability

Method used

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  • A method to identify whether Vibrio harveii is pathogenic
  • A method to identify whether Vibrio harveii is pathogenic
  • A method to identify whether Vibrio harveii is pathogenic

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1: Determination of detection standard and establishment of method

[0027] Vibrio harveyi strains from different sources were expanded and cultured by spectrophotometer and plate counting method, and then diluted with PBS solution to a concentration of 8×10 8 CFU / mL of bacterial liquid. 150-200 g of healthy large yellow croakers were selected for each strain and randomly divided into three test groups and a control group, with ten fish in each group. 100 microliters, 200 microliters, and 300 microliters were injected intramuscularly, and the control group was injected with the same dose of PBS solution. The pathogenic Vibrio harveii with a mortality rate of 100% within seven days is a non-pathogenic Vibrio harveyi with a lethal rate of less than 20%.

[0028] 2. Culture of pathogenic and non-pathogenic Vibrio harveyi

[0029] Pathogenic and non-pathogenic Vibrio harveyi were inoculated to seawater beef extract peptone solid culture, and cultured by streak...

Embodiment 2

[0051] Embodiment 2: detection of unknown Vibrio harveyi pathogenicity

[0052] 1. Cultivation of unknown Vibrio harveyi species

[0053] The Vibrio harveyi to be tested was inoculated with seawater beef extract peptone solid culture, and cultured by streaking. Incubate at 28°C for 24 hours. The medium formula is as follows: 1% peptone, 0.3% beef extract, 2% agar, and seawater is added to a certain value.

[0054] 2. Experiment of large yellow croaker infected by Vibrio harveyi

[0055] Using the spectrophotometer method and plate counting method, the Vibrio harveii to be tested after the expansion culture was diluted with PBS solution to a bacterial solution with a concentration of 8×108 CFU / mL. 150-200 g of healthy large yellow croakers were selected for each strain and randomly divided into three test groups and a control group, with ten fish in each group. 100 microliters, 200 microliters, and 300 microliters were injected intramuscularly, and the control group was inj...

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Abstract

The invention provides a method for identifying pathogenicity of Vibrio harveyi. A standard for detecting whether Vibrio harveyi is pathogenic is determined after metabolites of pathogenic Vibrio harveyi and non-pathogenic Vibrio harveyi are analyzed. Hence, the method is established on the basis of NMR technology based detection and analysis of characteristic metabolites of pathogenic Vibrio harveyi; the problem about identification of pathogenicity attributes of unknown Vibrio harveyi is solved. The experimental period is shortened, and the individual sensitivity difference of experimental animals and sensitivity of the experimental animals to the environment in the temporary breeding process are avoided, so that pathogenicity detection is more convenient and quicker.

Description

technical field [0001] The invention belongs to the technical field of pathogenicity identification of pathogenic bacteria, and in particular relates to a method for identifying whether Vibrio harveyi has pathogenicity. Background technique [0002] Vibrio harveyi (Vibrio harveyi) is a Gram-negative, luminescent marine bacterium that is widely distributed in the marine environment and can be divided into two types: pathogenic and non-pathogenic. Pathogenic Vibrio harveyi causes huge economic losses in aquaculture worldwide, especially in South America and Asia, where it can cause serious damage to marine fish, shrimp, shellfish, large yellow croaker, grouper, sea bream , perch, clams, prawns and other aquatic animal diseases, and can also cause food poisoning through the food chain, is a zoonotic pathogenic bacteria. At present, the detection methods of Vibrio harveyi mainly include physiological and biochemical methods, molecular biology and immunodiagnostic techniques, su...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N24/08
CPCG01N24/08
Inventor 张春丹苏秀榕叶央芳周君芦晨阳
Owner NINGBO UNIV
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