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Salmonella/Shigella/Staphylococcus aureus composite enriched medium and preparation method thereof

A Shigella and Salmonella technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of complex food matrix, complex ingredients, cumbersome preparation, etc., to achieve convenient preparation, simple ingredients, and fast proliferating effect

Inactive Publication Date: 2014-12-10
CAPITALBIO CORP +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the food matrix is ​​complex and the concentration of target bacteria is low. To meet the current technical detection requirements, a pre-enrichment step is still required to achieve high sensitivity.
At present, there are many kinds of pre-enrichment culture medium, but most of them are complicated in composition and cumbersome to prepare. Therefore, it is necessary to develop a medium with simple composition and convenient preparation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Weigh 5g of tryptone, 7.5g of yeast extract, 15g of sodium chloride, 3g of glucose, 1.5g of disodium hydrogen phosphate, 3g of mannitol, 1g of sodium pyruvate, 1g of glycine, and 1000ml of distilled water. L of NaOH was used to correct the pH value to 7.2, autoclaved at 121°C for 15 minutes, and stored at 4°C.

[0019] The cell concentration was 10 9 CFU / mL Salmonella, Shigella and Staphylococcus aureus were diluted 10 7 times, and then take 100 μL respectively, and insert into 10 mL prepared culture medium for Salmonella, Shigella and Staphylococcus aureus, so that the concentration of each bacteria in the culture medium is 1 CFU / mL, After culturing at 37°C for 16 hours, take 100 μL of the enrichment solution after the enrichment culture, properly dilute it and spread it on XLD agar and Baird-parker plates for isolation and culture. Count according to the characteristic colonies of Salmonella, Shigella and Staphylococcus aureus on the plate. The results are shown i...

Embodiment 2

[0024] Weigh 15g of tryptone, 2.5g of yeast extract, 5g of sodium chloride, 1g of glucose, 3.5g of disodium hydrogen phosphate, 1g of mannitol, 3g of sodium pyruvate, 3g of glycine, 1000ml of distilled water, stir and dissolve, and dissolve with 10 mol / L of NaOH was used to correct the pH value to 7.5, autoclaved at 121°C for 20min, and stored at 4°C.

[0025] The cell concentration was 10 9 CFU / mL Salmonella, Shigella and Staphylococcus aureus were diluted 10 7 times, and then take 100 μL respectively, and insert into 10 mL prepared culture medium for Salmonella, Shigella and Staphylococcus aureus, so that the concentration of each bacteria in the culture medium is 1 CFU / mL, After culturing at 37°C for 16 hours, take 100 μL of the enrichment solution after the enrichment culture, properly dilute it and spread it on XLD agar and Baird-parker plates for isolation and culture. Count according to the characteristic colonies of Salmonella, Shigella and Staphylococcus aureus on...

Embodiment 3

[0030] Weigh 10g of tryptone, 5g of yeast extract, 10g of sodium chloride, 2g of glucose, 2.5g of disodium hydrogen phosphate, 2g of mannitol, 2g of sodium pyruvate, 2g of glycine, 1000ml of distilled water, stir to dissolve, and add 10 mol / L Correct the pH value to 7.4 with NaOH, autoclave at 121°C for 20min, and store at 4°C.

[0031] The cell concentration was 10 9 CFU / mL Salmonella, Shigella and Staphylococcus aureus were diluted 10 7 times, and then take 100 μL respectively, and insert into 10 mL prepared culture medium for Salmonella, Shigella and Staphylococcus aureus, so that the concentration of each bacteria in the culture medium is 1 CFU / mL, After culturing at 37°C for 16 hours, take 100 μL of the enrichment solution after the enrichment culture, properly dilute it and spread it on XLD agar and Baird-parker plates for isolation and culture. Count according to the characteristic colonies of Salmonella, Shigella and Staphylococcus aureus on the plate. The results ...

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Abstract

The invention discloses a Salmonella / Shigella / Staphylococcus aureus composite enriched medium and a preparation method thereof. The medium comprises the following components in parts by mass: 5-15 parts of tryptone, 2.5-7.5 parts of yeast extract, 5-15 parts of sodium chloride, 1-3 parts of glucose, 1.5-3 parts of disodium hydrogen phosphate, 1-3 parts of mannitol, 1-3 parts of sodium pyruvate, 1-3 parts of glycine and 1000 parts of distilled water. The pH value is 7.2-7.5. The preparation method comprises the following steps: adding all the components to the distilled water, dissolving by stirring and autoclaving. The Salmonella, Shigella and Staphylococcus aureus are main pathogens in need of detection in national standard of food sanitation. The composite enriched medium can be utilized to implement constant-speed and high-speed proliferation of the Salmonella, Shigella and Staphylococcus aureus. After proliferation for 16 hours, the composite enriched medium can be directly used for high-flux detection of multiplex PCR (polymerase chain reaction), gene chips, microfluidic chips and the like to screen the three pathogens.

Description

technical field [0001] The invention belongs to a pre-enrichment culture medium for pathogenic bacteria, in particular to a culture medium for compound enrichment of Salmonella, Shigella and Staphylococcus aureus at the same time and a preparation method thereof. Background technique [0002] Bacterial microorganisms are the main cause of foodborne diseases in my country, seriously endangering people's health and causing heavy economic losses, and have become one of the more prominent public health problems at present. Salmonella, Shigella and Staphylococcus aureus are the main pathogenic bacteria of foodborne diseases, and they are also important indicators for food safety risk monitoring in my country. China's foodborne disease statistics report shows that more than 30% of the related cases are related to these three bacteria, and Salmonella ranks first. Today, people pay more and more attention to food safety issues, and it is imminent to establish a simultaneous detecti...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/42C12R1/445C12R1/01
Inventor 张岩刘铭李云葛少林邢婉丽
Owner CAPITALBIO CORP
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