A kind of in vivo drug resistance induction model of Pseudomonas aeruginosa and its construction method and application
A technology of Pseudomonas aeruginosa and a construction method, which is applied to the in vivo drug resistance induction model of Pseudomonas aeruginosa animals and the field of its construction, and can solve the problem of not being able to simulate the development law of Pseudomonas aeruginosa drug resistance and model evaluation. Inaccurate standards, lack of Pseudomonas aeruginosa models, etc.
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Embodiment 1
[0072] This embodiment provides a method for constructing a drug resistance induction model in PA animals, comprising the following steps:
[0073](1) Take 200 healthy ICR mice, half male and half female, weighing 14±1g, and randomly group them into groups. The induction modeling period is set to 3 days, 5 days, 7 days, and 9 days respectively, and a blank is set for each induction modeling period. Control group, non-induced drug resistance model group A1, non-induced drug resistance model administration group A2, induced drug resistance model group B1 and induced drug resistance model administration group B2, a total of 20 groups (see Table 2 below), each group 10, spare; the non-induced drug resistance model group A1 and the non-induced drug resistance model administration group A2 are collectively referred to as group A, and the induced drug resistance model group B1 and the induced drug resistance model administration group B2 are collectively referred to as Group B;
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Embodiment 2
[0101] In this example, on the basis of Example 1, the expression level of the efflux pump gene mexC in the mouse lung tissue is used to verify whether the drug resistance induction model in the PA animal is successfully constructed. The method for expressing the amount of mexC gene comprises the steps:
[0102] 1) Collect the lung tissue of each group of mice described in Example 1 as a sample and place it in a mortar, add liquid nitrogen, grind it finely, collect it in a 1.5ml centrifuge tube, add 1ml Trizol to each sample, and extract RNA; Flick the bottom of the EP tube with 1ml TrizolReagent to resuspend the mixed sample, place at room temperature for 20min, 4°C, 12000rpm, centrifuge for 10min; transfer the clarified supernatant into a new EP tube, add 0.2ml chloroform, and cap tightly Cover the tube, shake vigorously for 15s, incubate at room temperature for 2-3min, 4°C, 12000rpm, centrifuge for 15min; transfer the supernatant to a new EP tube, add 0.5ml isopropanol, mix...
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