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Building method for mouse model systemically infected with pseudomonas aeruginosa

A Pseudomonas aeruginosa and mouse model technology, applied in the biological field, can solve the problems of not being able to represent the infection situation, and achieve the effects of easy control of infection conditions, high infection rate, and low bacterial concentration

Inactive Publication Date: 2016-07-20
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no literature report on the systemic infection model caused by Pseudomonas aeruginosa
In addition, the Pseudomonas aeruginosa strain used in the reported infection model is the standard strain PA01 from the United States, which cannot represent the infection situation of the domestic popular PA strains

Method used

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  • Building method for mouse model systemically infected with pseudomonas aeruginosa
  • Building method for mouse model systemically infected with pseudomonas aeruginosa
  • Building method for mouse model systemically infected with pseudomonas aeruginosa

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: preparation of Pseudomonas aeruginosa bacterium liquid

[0034] Take the frozen Pseudomonas aeruginosa clinical strain XN-1, revive it with LB nutrient agar plate, and culture it aerobically at 37°C overnight. Pick a single colony and inoculate 20ml of LB liquid medium, and incubate with aerobic shaking at 180rpm at 37°C for 15h, then inoculate 0.2mL into 20ml of LB liquid medium, and inoculate at 37°C with aerobic shaking at 230rpm for 6h. The cells were collected by centrifugation at 6000g, washed twice with normal saline, and then resuspended with normal saline. Adjust the bacterial concentration to OD 600 The value is 0.8, and the bacterial concentration measured by plate counting method is 3.0×10 9 CFU / ml.

Embodiment 2

[0035] Example 2: Establishment of mouse tail vein injection scheme

[0036] In the infection experiment of the whole body model of Pseudomonas aeruginosa, the control of the intravenous dose is the guarantee of the stability of the model. In order to better control the intravenous dose and prevent the bacterial liquid from entering the soft tissue around the blood vessel, the following measures have been taken: (1 ) Irradiating the mouse with infrared light for 3 minutes, it can be seen that the veins on the left and right sides of the tail of the mouse are obviously filled and expanded; (2) Fix the mouse with a mouse fixer, and insert the needle at a distance of 2 to 3 cm from the tail of the mouse at less than 30 degrees; (3) Disposable sterile insulin syringes are used, with short and thin needles; (4) There will be a sense of void when the needle enters the blood vessel, and there is no resistance when pushing, and blood will return when the needle is pulled out; (5) In th...

Embodiment 3

[0037] Example 3: Determination of Infection Dose

[0038] Physiological saline was used to adjust the PAXN-1 bacterial solution obtained in Example 1 to six different concentrations, and the experimental animal female BALB / c mice were randomly divided into 7 groups, and were infected by tail vein injection. The infection dose of each mouse was 100 μL, and the same dose of normal saline (NS) was used as a blank control. The grouping and infection status of the animals are shown in Table 1. After the infection, the death of the mice was observed every other day, and the observation period was 7 days. After the observation period, the remaining animals were treated with CO 2 Euthanized by inhalation.

[0039] Table 1: Determination of Infection Dose in Pseudomonas aeruginosa Systemic Model

[0040]

[0041]

[0042] When adopting the Pseudomonas aeruginosa bacterial strain PAXN-1 infection BALB / c mouse of different concentrations (2-fold gradient dilution), through 7 da...

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Abstract

The invention relates to a building method for a mouse model systemically infected with pseudomonas aeruginosa. The building method comprises the following steps that 1, a mouse is irradiated by an infrared lamp for 3 minutes, and the vein vessels on the two sides of the tail of the mouse are obviously filled and dilated; 2, pseudomonas aeruginosa is intravenously injected into the tail of the mouse. According to the method, the mouse can be effectively infected, and the model systemically infected with the pseudomonas aeruginosa is successfully built. The infected mouse model is stable and can be used for vaccine evaluation and protective mechanism research.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for establishing a mouse model of systemic infection by Pseudomonas aeruginosa. Background technique [0002] Pseudomonas aeruginosa (Pseudomonas aeruginosa, PA), commonly known as Pseudomonas aeruginosa, belongs to the genus Pseudomonas in non-fermenting bacteria. It is ubiquitous and is one of the most common opportunistic pathogens in clinical practice. At present, the bacterium has become one of the pathogenic bacteria with the highest isolation rate in ICU wards, burns, war wound infections, and mechanical ventilation-associated pneumonia (VAP) worldwide. PA infection can occur in any part and tissue of the human body, such as burns or trauma, middle ear, cornea, urethra, and respiratory tract, etc. It can also cause systemic infections such as endocarditis, gastroenteritis, empyema, and even sepsis, and death from systemic infection The rate exceeds 20%. In addition...

Claims

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Application Information

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IPC IPC(8): A01K67/02A61K35/74A61K49/00
CPCA01K67/02A61K35/74A61K49/0008
Inventor 章金勇顾江杨峰邹全明彭六生赵莉群敬海明王逸麟秦溢
Owner ARMY MEDICAL UNIV
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