A kind of establishment method of Staphylococcus aureus pneumonia model

A method for establishing a staphylococcus infection technology, applied in the field of building an animal model of Staphylococcus aureus pneumonia, can solve the problems of death of experimental animals, lack of susceptible models, damage to animal organs, etc., and achieves high infection rate and easy control of infection conditions. , the effect of infection stabilization

Active Publication Date: 2016-10-12
CHENGDU OLYMVAX BIOPHARM +1
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Problems solved by technology

But these methods still have big problems, which are mainly reflected in the following aspects: 1. Due to the structure of the connection between the nasal cavity and the oral cavity, the bacteria liquid directly dripped into the mouse without anesthesia easily enters the mouth and cannot accurately control the bacteria liquid entering the lungs, that is, it cannot be accurately controlled. Controlling the dose of infection leads to unstable model construction; 2. Although the method of inserting the trachea can guarantee the dose of infection, as an invasive method, it is easy to cause damage to animal organs or even death of experimental animals during the operation; 3. There are too many species to choose, and there is a lack of a clear susceptibility model; 4. The colonization of bacterial infection is unstable, which may easily cause the instability of the model, and the stability of the infection dose is closely related to the success rate of modeling

Method used

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  • A kind of establishment method of Staphylococcus aureus pneumonia model
  • A kind of establishment method of Staphylococcus aureus pneumonia model
  • A kind of establishment method of Staphylococcus aureus pneumonia model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: preparation of Staphylococcus aureus bacterial liquid

[0027] Take the frozen MRSA252, revive it with MH agar plate, and culture it aerobically at 37°C overnight. Pick a single colony and inoculate 1000ml of MH liquid medium, culture with aerobic shaking at 210rpm for 7 hours at 37°C, collect the bacteria by centrifugation at 4700rmp, wash twice with normal saline and resuspend the bacteria with normal saline. The OD of the bacterial solution was measured by a spectrophotometer 600 value, and according to 1OD=2×10 9 CFU / ml was converted into bacterial concentration.

Embodiment 2

[0028] Example 2: Establishment of mouse anesthesia and nasal drop scheme

[0029] In the infection experiment of the Staphylococcus aureus 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

[0030] Embodiment 3: Preliminary groping for the dose of infection

[0031] The MRSA252 bacterial solution obtained in Example 1 was adjusted to five different concentrations with physiological saline, and the experimental animal female C57BL / 6J mice were randomly divided into 5 groups. After the mice were anesthetized with isoflurane, they were infected by nasal drops. The infection dose of each mouse was 50 μL, and the same dose of normal saline (NS) was used as the 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 10 days. After the observation period, the remaining animals were treated with CO 2 Euthanized by inhalation.

[0032] Table 1: Preliminary exploration of the infection dose of the Staphylococcus aureus pneumonia model

[0033] SA strain

[0034] When C57BL / 6J mice were initially infected with different concentrat...

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Abstract

The invention belongs to the field of biotechnologies, and provides a building method of an SA pneumonia animal model. According to the building method, an infection condition is controlled easily, the infection success rate is high, infection stability is good, and an obvious symptom of pneumonia of a mouse can be caused.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the establishment of an animal model, more specifically, to a method for establishing an animal model of Staphylococcus aureus pneumonia. Background technique [0002] Staphylococcus aureus (Staphylococcus aureus, SA), hereinafter referred to as Staphylococcus aureus, has another name of "flesh-loving bacteria". As a representative of Gram-positive bacteria, it is an important pathogenic bacteria causing nosocomial infection and community infection. Infection is characterized by acute, purulent inflammation. Local infection can cause purulent infection of the skin and soft tissues, which does not heal after a long time; systemic infection can lead to osteomyelitis, septic arthritis, endocarditis, pneumonia, and sepsis And other serious infections and complications, the mortality rate is as high as 20%. At the same time, the exotoxin of Staphylococcus aureus can also cause systemic fa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61D7/00A01K67/027
Inventor 王逸麟曾浩邹全明章金勇敬海明董衍东解庆华
Owner CHENGDU OLYMVAX BIOPHARM
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