Culture medium for composite enrichment of staphylococcus aureus, salmonella and listeria monocytogenes and preparation method

A technology for Listeria monocytogenes and Staphylococcus, which is applied in the field of pre-enrichment culture medium for pathogenic bacteria, can solve the problems of increasing the workload of detection, etc., and achieve the effects of rapid proliferation, convenient preparation, and good co-enrichment effect

Active Publication Date: 2020-08-07
JILIN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to overcome the problems in the prior art that different enrichment media are often used for cultivating different pathogenic bacteria, which increases the workload of detection, the present invention provides a method for Staphylococcus aureus, Salmonella and monoenrichment The culture medium and preparation method for the compound enrichment of Listeria are suitable for high-throughput detection equipment such as multiplex PCR, and help to realize the simultaneous and rapid detection of three pathogenic bacteria

Method used

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  • Culture medium for composite enrichment of staphylococcus aureus, salmonella and listeria monocytogenes and preparation method
  • Culture medium for composite enrichment of staphylococcus aureus, salmonella and listeria monocytogenes and preparation method
  • Culture medium for composite enrichment of staphylococcus aureus, salmonella and listeria monocytogenes and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Identification of the monoenrichment effect of SSL medium

[0033] A kind of culture medium for the compound enrichment of Staphylococcus aureus, Salmonella and Listeria monocytogenes of this embodiment, in terms of mass parts, its formula consists of: 17.0 parts of tryptone, 3.0 parts of peptone, yeast extract 6.0 parts, 2.5 parts of dipotassium hydrogen phosphate, 9.6 parts of disodium hydrogen phosphate, 1.35 parts of potassium dihydrogen phosphate, 15.0 parts of peptone, 20.0 parts of beef powder, 10.0 parts of soybean peptone, 0.3 parts of cycloheximide, 3.5 parts of mannitol, 4.5 parts of sodium pyruvate, 0.00115 parts of fosfomycin sodium, 1.0 parts of lithium chloride, 1000 parts of distilled water, pH The value is 7.2.

[0034] A kind of preparation method for the compound enrichment culture medium of Staphylococcus aureus, Salmonella and Listeria monocytogenes of this embodiment specifically comprises the following steps:

[0035] (1), mix trypto...

Embodiment 2

[0039] Example 2: Identification of the co-enrichment effect of SSL medium

[0040] A certain amount of Staphylococcus aureus, Salmonella, and Listeria monocytogenes were added to the SSL medium, so that the initial concentration of the three target bacteria was 100 CFU / mL. After inoculation, vibrate culture at 37°C and 200rpm, take cultures at 6h, 12h, 16h, 20h, and 24h for plate counting, and the counting media for Staphylococcus aureus, Salmonella, and Listeria monocytogenes are respectively B-P medium, Salmonella chromogenic medium, and PALCAM medium.

[0041] According to the colony counting results at each time point, the figure 2 From the growth curve shown, it can be seen from the figure that 0-12 h is the stage of rapid bacterial growth. After 12 hours of cultivation, the concentrations of the three bacteria were basically at 10 8 More than CFU / mL (Staphylococcus aureus 0.83×10 8 CFU / mL, Salmonella 1.15×10 9 CFU / mL, Listeria monocytogenes 1.25 ×10 8 CFU / mL), th...

Embodiment 3

[0042] Embodiment 3: the inhibitory effect of SSL medium to non-target bacteria

[0043] A certain amount of non-target bacteria was added to SSL medium and tryptone soy broth (TSB) medium separately, with an initial concentration of 100 CFU / mL, and then cultured with shaking at 37°C and 200rpm. The cultures of 6h, 12h, 16h, 20h, and 24h were taken to measure the McFarland turbidity value, and the results are shown in Table 4. Compared with the growth in TSB medium, it can be seen that the growth of non-target bacteria Bacillus subtilis, Yersinia enterocolitica and Vibrio parahaemolyticus in SSL medium was significantly inhibited.

[0044] Table 4 McFarland turbidity values ​​at different time points when non-target bacteria were cultured in SSL and TSB medium

[0045]

[0046]

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Abstract

The invention discloses a culture medium for composite enrichment of staphylococcus aureus, salmonella and listeria monocytogenes and a preparation method. The invention belongs to the technical fieldof pre-enrichment culture media of pathogenic bacteria. The culture medium comprises, by mass, 15.0 to 19.0 parts of tryptone, 2.7 to 3.3 parts of peptone, 5.4 to 6.6 parts of yeast extract, 2.25 to2.75 parts of dipotassium hydrogen phosphate, 8.6 to 10.6 parts of disodium hydrogen phosphate, 1.22 to 1.48 parts of potassium dihydrogen phosphate, 13.0 to 17.0 parts of peptone, 18.0 to 21.0 partsof beef powder, 8.0 to 12.0 parts of soy peptone, 0.2 to 0.4 part of actinone, 2.0 to 4.0 parts of mannitol, 4.0 to 5.0 parts of sodium pyruvate, 0.00078 to 0.00156 part of fosfomycin sodium, 0.75 to1.1 parts of lithium chloride and 1000 parts of distilled water, wherein the pH value is 6.8 to 7.4. The single enrichment effect of the composite enrichment culture medium is superior to that of respective selective enrichment liquid, and growth of non-target bacteria can be well inhibited. Under the condition that non-target bacteria exist, the culture medium can also achieve constant-speed andrapid proliferation of the three target bacteria.

Description

technical field [0001] The invention belongs to the technical field of pre-enrichment medium for pathogenic bacteria, and in particular relates to a medium for compound enrichment of Staphylococcus aureus, Salmonella and Listeria monocytogenes and a preparation method thereof. Background technique [0002] In recent years, food safety has attracted more and more attention at home and abroad, and food-borne pathogens are one of the most important factors threatening food safety. According to the statistics of the World Health Organization, about 600 million people are sick and more than 400,000 people die every year due to food-borne pathogens in the world. How to effectively monitor the pathogenic bacteria contamination of food in the links of planting, processing, circulation and consumption has become an important practical problem that people urgently need to solve. [0003] Salmonella, Staphylococcus aureus and Listeria monocytogenes are the main pathogenic bacteria com...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/445C12R1/42C12R1/01
CPCC12N1/20Y02A50/30
Inventor 张晓光藏程琳
Owner JILIN UNIV
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