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Method for establishing pseudomonas aeruginosa infectious pneumonia animal model

A Pseudomonas aeruginosa, animal model technology, applied in pharmaceutical formulations, compound screening/testing, medical raw materials derived from bacteria, etc., can solve the problem that cannot represent the prevalence of PA strains, it is difficult to carry out experiments in large quantities, and the death of experimental animals and other problems to achieve the effect of avoiding the death of experimental animals, being representative, and easy to infect conditions

Active Publication Date: 2016-07-13
ARMY MEDICAL UNIV
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  • Application Information

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Problems solved by technology

However, there are still big problems in these methods, which are mainly reflected in the following aspects: 1. Although the method of incision of the trachea can guarantee the infection dose, as an invasive method, it is easy to cause damage to the animal organs during the operation and even lead to experimental failure. The animal died; 2. The operation is complicated, and each mouse needs to be operated on, so it is difficult to carry out experiments in large quantities; 3. The Pseudomonas aeruginosa strain used in the reported infection model is the standard strain PA01 from the United States, which cannot represent Epidemiological situation of popular PA strains in China

Method used

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  • Method for establishing pseudomonas aeruginosa infectious pneumonia animal model
  • Method for establishing pseudomonas aeruginosa infectious pneumonia animal model
  • Method for establishing pseudomonas aeruginosa infectious pneumonia animal model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: preparation of Pseudomonas aeruginosa bacterium liquid

[0039] 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 for 15 hours at 37°C, then inoculate 0.2mL into 20mL of LB liquid medium, and culture with aerobic shaking at 230rpm for 6h at 37°C. The cells were collected by centrifugation at 6000g, washed twice with normal saline, and then resuspended with normal saline. The OD of the bacterial solution was measured by a spectrophotometer 600 Value, and adjust the OD value to 1.0 with normal saline, through serial dilution, and count on LB plates to obtain Pseudomonas aeruginosa at OD=1.0 when it is 6.0×10 9 CFU / ml.

Embodiment 2

[0040] Example 2: Establishment of mouse anesthesia and nasal drop scheme

[0041]In the infection experiment of the Pseudomonas aeruginosa pneumonia model, in order to ensure the smooth progress of the nasal drip process, the mice were first anesthetized with isoflurane. Anesthesia method: Inhalation induction anesthesia with 5% isoflurane using oxygen as the transport carrier, and maintain anesthesia with 3% concentration after the mice enter the surgical deep anesthesia period, this method can achieve better anesthesia effect. The control of the nasal drop dose is the guarantee of the stability of the model. In order to better control the nasal drop dose and prevent the bacterial liquid from entering the mouth and generating air bubbles during the nasal drop process, the following measures have been taken: (1) Since the nasopharynx has a physiological The more the head is tilted, the easier it is for nasal drops to enter the mouth. Therefore, during the nasal dripping proce...

Embodiment 3

[0042] Example 3: Determination of Infection Dose

[0043] Physiological saline was used to adjust the PAXN-1 bacterial solution obtained in Example 1 to five different concentrations, and the experimental animal female BALB / c mice were randomly divided into 5 groups, and the mice were anesthetized with isoflurane and infected by nasal drops , the infection dose for each mouse was 20 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.

[0044] Table 1: Determination of Infection Dose in Pseudomonas Aeruginosa Pneumonia Model

[0045] Group Nasal drip Dose (CFU) quantity Concentration (CFU / ml) Dropping amount (μl) 1 PAXN-1 6.0×10 8 10 3.0×10 10 ...

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Abstract

The invention relates to a method for establishing a pseudomonas aeruginosa infectious pneumonia animal model. The method comprises the following steps: 1) deeply anesthetizing a mouse by adopting isoflurane; and 2) dripping nose of the mouse with pseudomonas aeruginosa, so that a pseudomonas aeruginosa infectious pneumonia model is established. The method provided by the invention has the advantages that the mouse can be effectively infected, and the pseudomonas aeruginosa pneumonia model is successfully established; and the mouse model after infection is stable and can be applied to evaluation of vaccines and research of protective mechanisms.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for establishing an animal model of Pseudomonas aeruginosa infection pneumonia. 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., and can also cause systemic infections such as endocarditis, gastroenteritis, pneumonia, and even sepsis, and cause systemic infection mortality More than 20%. In additi...

Claims

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Application Information

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IPC IPC(8): A61K49/00A61K35/74
CPCA61K35/74A61K49/0008
Inventor 顾江杨峰章金勇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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