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Composite enrichment medium for Escherichia coli, Salmonella and Listeria monocytogenes

A technology for Listeria monocytogenes and Salmonella, applied in the direction of bacteria, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of limited application and poor bacterial enrichment effect, and achieve rapid proliferation and convenient preparation Effect

Inactive Publication Date: 2019-07-05
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Since each pathogenic bacteria has its own growth habits and nutritional requirements, different enrichment media are often required for different pathogenic bacteria, which largely limits the biosensors such as multiplex PCR and multichannel SPR. Application of high-throughput detection technologies such as multi-channel BLI biosensors in actual detection
At present, there are some studies on co-enrichment culture at home and abroad, but there are very few co-enrichment culture media that can be used for E. coli O157:H7, Salmonella and Listeria monocytogenes, and the enrichment effect is not very good

Method used

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  • Composite enrichment medium for Escherichia coli, Salmonella and Listeria monocytogenes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Weigh 17.0 parts of tryptone, 3.0 parts of peptone, 6.0 parts of yeast extract, 10.0 parts of sodium chloride, 2.5 parts of dipotassium hydrogen phosphate, 9.6 parts of disodium hydrogen phosphate, 1.35 parts of potassium dihydrogen phosphate, and 0.00125 parts of acridine yellow , 5.0 parts of magnesium chloride, 4.0 parts of glucose, 5.0 parts of sodium pyruvate, distilled water to 1000 parts, heated to dissolve, cooled to room temperature, corrected to pH 7.2, autoclaved at 121°C for 20 minutes, and stored at 4°C.

[0018] The overnight culture of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes was serially diluted with PBS buffer, and 10 7 Each 400 μL of the two-fold dilution was added to 225 mL of co-enrichment culture medium. Take 25mL of sterile water and put it into the inoculated co-enrichment medium, mix well and incubate at 37°C for 16h. Take 100 μl of the enrichment solution after the enrichment culture, and spread it on Sorbitol MacConkey ...

Embodiment 2

[0023] Preparation of the culture medium for compound enrichment of Escherichia coli O157:H7, Salmonella and Listeria monocytogenes: refer to Example 1.

[0024] The overnight culture of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes was serially diluted with PBS buffer, and 10 7 Each 400 μL of the two-fold dilution was added to 225 mL of co-enrichment culture medium. Take 25mL of sterile water and put it into the inoculated co-enrichment medium, mix well and incubate at 37°C for 24h. Take 100 μl of the enrichment solution after the enrichment culture, and spread it on Sorbitol MacConkey agar medium, Salmonella selective agar medium, Listeria selective agar medium plate respectively after appropriate dilution for culture counting. The counting results are shown in Table 2.

[0025] Table 2 24-hour bacterial enrichment effect of compound bacterial enrichment medium

[0026] strain name Inoculum concentration (CFU / mL) Bacteria concentration afte...

Embodiment 3

[0029] Preparation of the culture medium for compound enrichment of Escherichia coli O157:H7, Salmonella and Listeria monocytogenes: refer to Example 1.

[0030] The overnight culture of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes was serially diluted with PBS buffer, and 10 7 Inoculate 400 μL of each double dilution on 25 g of chicken, add 225 mL of co-enrichment culture medium, homogenize for 1-2 minutes with a beating homogenizer, and then incubate at 37°C for 24 hours. Take 100 μl of the enrichment solution after the enrichment culture, and spread it on Sorbitol MacConkey agar medium, Salmonella selective agar medium, Listeria selective agar medium plate respectively after appropriate dilution for culture counting. The counting results are shown in Table 3.

[0031] Table 3 The enrichment effect of compound enrichment medium applied to actual samples

[0032]

[0033] It can be seen from Table 3 that after 24 hours of enrichment, the concentration...

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Abstract

The invention discloses a composite enrichment medium for Escherichia coli, Salmonella and Listeria monocytogenes and a preparation method thereof. The composite enrichment medium for Escherichia coli, Salmonella and Listeria monocytogenes comprises 16.0-18.0 parts of tryptone, 2.0-4.0 parts of peptone, 5.0-7.0 parts of yeast extract, 10.0-20.0 parts of sodium chloride, 2.4-2.6 parts of dipotassium phosphate, 9.5-9.7 parts of disodium hydrogen phosphate, 1.34-1.36 parts of monopotassium phosphate, 0.005-0.020 part of acriflavine, 2.0-8.0 pars of magnesium chloride, 1.0-4.0 parts of glucose, 1.0-4.0 parts of sodium pyruvate and 1000 parts of distilled water, and has pH of 6.8-7.4. The composite enrichment medium herein allows three target pathogens to be enriched at a high, constant speed;after 16-24 hours of enrichment, quick detection of the three target pathogens can be implemented to provide a diagnostic report through high-flux detection devices, such as multiple PCR (polymerase chain reaction), multi-channel SPR (surface plasma resonance) biosensors, and multi-channel BLI (biolayer interferometry) biosensors.

Description

technical field [0001] The invention belongs to a pre-enrichment medium for pathogenic bacteria, in particular to a medium for compound enrichment of Escherichia coli O157:H7, Salmonella and Listeria monocytogenes at the same time and a preparation method thereof. Background technique [0002] Foodborne illness is a worldwide public health problem. The economic loss caused by foodborne diseases in the United States is as high as 110 billion yuan every year. Food-borne pathogens are the most important factor causing food-borne diseases. How to effectively monitor microbial contamination in areas such as food production and circulation has become a realistic problem that people must face. [0003] Escherichia coli O157:H7, Salmonella and Listeria monocytogenes are three common food-borne pathogens that must be detected in my country's national food safety standards. The detection of these foodborne pathogens, whether it is the classic traditional culture detection method or...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/19C12R1/42C12R1/01
CPCC12N1/20
Inventor 张鸣镝藏程琳张晓光吴平平张佐敏谢楠楠杜煜
Owner JILIN UNIV
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