Screening culture medium applicable to extensively drug-resistant pseudomonas aeruginosa and preparation method

A technique for screening culture medium for Pseudomonas aeruginosa, which is applied in the field of microorganisms to achieve the effects of shortening detection time, promoting growth and secretion of aeruginosa, and shortening screening time

Active Publication Date: 2017-07-21
廊坊恒益生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The object of the present invention is to provide a kind of screening culture medium and preparation method applicable to extensively drug-resistant Pseudomonas aeruginosa, thereby solving the aforementioned problems existing in the prior art

Method used

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  • Screening culture medium applicable to extensively drug-resistant pseudomonas aeruginosa and preparation method
  • Screening culture medium applicable to extensively drug-resistant pseudomonas aeruginosa and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: the preparation of the nutrient substance that promotes the growth of Pseudomonas aeruginosa

[0039] 1. Pretreatment of animal blood: Purchase 1000ml of pig blood at the farmer’s market, take 100ml and add 900ml of distilled water, use SKG 1246 wall-breaking food processor to stir and break the clot, and obtain blood mixture, the speed is 1200 rpm, and the time is 3 minute.

[0040] 2. Adjust the pH value: use 1mol / L sodium hydroxide (pH: 14) to adjust the pH value of the blood mixture to 8.5 to obtain the blood raw material solution;

[0041] 3. High-temperature and high-pressure hydrolysis: Put the blood raw material liquid into a high-temperature and high-pressure reaction kettle, and carry out high-temperature and high-pressure hydrolysis reaction while stirring under the conditions of temperature 195°C, atmospheric pressure 2.1 atmospheres, and stirring speed of 100 rpm. For 1h, the hydrolyzate was obtained, and the high-temperature and high-pressur...

Embodiment 2

[0044] Embodiment 2: NAC agar plate and the NAC agar plate (without adding antibiotic combination) comparative test of adding the nutrient substance that promotes the growth of Pseudomonas aeruginosa

[0045] 1. Prepare NAC agar plate

[0046]Weigh 20g of peptone, 0.3g of dipotassium hydrogen phosphate, 0.2g of magnesium sulfate, 0.2g of cetrimethylene bromide, 0.015g of nalidixic acid, and 15g of agar, dissolve them in 1L of distilled water, mix well, and autoclave at 121°C for 25min Afterwards, it was poured onto a sterile plate and cooled at room temperature to obtain a NAC agar plate, which was set as the control group.

[0047] 2. NAC agar plates for preparing nutrients that promote the growth of Pseudomonas aeruginosa (without adding antibiotic combinations)

[0048] Weigh peptone 20g, dipotassium hydrogen phosphate 0.3g, magnesium sulfate 0.2g, bromide trimethylammonium 0.2g, nalidixic acid 0.015g, agar 15g, add 50ml filtrate and 950ml distilled water in Example 1, mix...

Embodiment 3

[0053] Example 3: NAC agar plate and the screening method based on the screening medium suitable for extensively drug-resistant Pseudomonas aeruginosa and its screening medium comparison test

[0054] 1. Prepare NAC agar plate

[0055] Weigh 20g of peptone, 0.3g of dipotassium hydrogen phosphate, 0.2g of magnesium sulfate, 0.2g of cetrimethylene bromide, 0.015g of nalidixic acid, and 15g of agar, dissolve them in 1L of distilled water, mix well, and autoclave at 121°C for 25min Afterwards, it was poured onto a sterile plate and cooled at room temperature to obtain a NAC agar plate, which was set as the control group.

[0056] 2. Preparation of agar plates for selective isolation and cultivation of extensively drug-resistant Pseudomonas aeruginosa (add antibiotic combination)

[0057] (1) Weigh 20g of peptone, 0.3g of dipotassium hydrogen phosphate, 0.2g of magnesium sulfate, 0.2g of cetrimethylene bromide, 0.015g of nalidixic acid, and 15g of agar, add 100mL of additive solut...

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Abstract

The invention discloses a screening culture medium applicable to extensively drug-resistant pseudomonas aeruginosa and a preparation method, and relates to the technical field of microorganism. The screening culture medium comprises a basal culture medium, nutrient substance promoting the growth of the pseudomonas aeruginosa, antibiotic used for the detection of the multidrug resistant pseudomonas aeruginosa and distilled water. The method comprises the following steps: preparing the nutrient substance promoting the growth of the pseudomonas aeruginosa, adding the nutrient substance into the basal culture medium, and performing autoclaving, so as to obtain an initial culture medium; adding the antibiotic used for the detection of the multidrug resistant pseudomonas aeruginosa into the initial culture medium at a room temperature to 60 DEG C after sterilization, performing uniformly mixing, separately injecting the initial culture medium into different holes of a layout plate, cooling at a room temperature, so as to obtain the screening culture medium applicable to the extensively drug-resistant pseudomonas aeruginosa. The screening culture medium has the advantages that the detection time is reduced, the sensitivity is high and the preparation method is simple and convenient.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a screening medium suitable for extensively drug-resistant Pseudomonas aeruginosa and a preparation method. Background technique [0002] Pseudomonas aeruginosa is a common opportunistic pathogen that can cause purulent lesions. After infection, the pus and exudate are green, so it is also known as Pseudomonas aeruginosa. It is one of the clinically important opportunistic pathogens. one. [0003] In clinical sample testing, highly suspicious colonies are often selected from the numerous colonies obtained from the test samples by using blood agar culture medium for isolation and culture, and after confirming that the cultured colonies are free from contamination by foreign bacteria, the cultured colonies System identification and drug susceptibility experiments were performed. The detection time of this traditional detection method usually reaches 72 hours or more, which...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12N1/38C12R1/385
CPCC12N1/38C12Q1/045
Inventor 宋征奇杨学敏朱克先谭谨刘丽霞刘红霞马晓玉
Owner 廊坊恒益生物技术有限公司
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