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Composite enrichment medium for five bacteria and preparation method for composite enrichment medium

A culture medium and formula technology, applied in the field of pre-enrichment medium, can solve the problems that are not suitable for Salmonella, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Vibrio parahaemolyticus

Active 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

In addition, there are general media such as UPB (Universal Pre-enrichment Broth), BPW (Buffered Peptone Water), and NB (Nutrient Broth), which can multiply a variety of bacteria at the same time, but they are not suitable for Salmonella and Staphylococcus aureus. , Escherichia coli, Listeria monocytogenes and Vibrio parahaemolyticus, five pathogenic bacteria with simple components and easy preparation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Weigh 5g of bovine brain extract powder, 15g of bovine heart extract powder, 5g of peptone, 2.5g of sodium chloride, 3g of glucose, 1.5g of disodium hydrogen phosphate, 1g of mannitol, 1g of sodium pyruvate, 3g of glycine, 1g of escin, 1000ml of distilled water, stir to dissolve, adjust the pH value to 7.2 with 10 mol / L NaOH solution, autoclave at 121°C for 15min, and store at 4°C.

[0016] The cell concentration was 10 9 CFU / mL Salmonella, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Vibrio parahaemolyticus were diluted 10 7 times, and then take 100 μL respectively, and add 10 mL of the prepared compound enrichment medium for Salmonella, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Vibrio parahaemolyticus to make the medium The concentration of each bacterium in the medium was 1 CFU / mL, cultured at 37°C for 16 hours, took 100 μL of the enrichment solution after the enrichment culture, and diluted it appropriately and appl...

Embodiment 2

[0021] Weigh bovine brain extract powder 10g, beef heart extract powder 5g, peptone 15g, sodium chloride 7.5g, glucose 1g, disodium hydrogen phosphate 3.5g, mannitol 3g, sodium pyruvate 3g, glycine 1g, escin 3g, 1000ml of distilled water, stir to dissolve, adjust the pH value to 7.5 with 10 mol / L NaOH solution, autoclave at 121°C for 17 min, and store at 4°C.

[0022] The cell concentration was 10 9 CFU / mL Salmonella, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Vibrio parahaemolyticus were diluted 10 7 times, and then take 100 μL respectively, and add 10 mL of the prepared compound enrichment medium for Salmonella, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Vibrio parahaemolyticus to make the medium The concentration of each bacterium in the medium was 1 CFU / mL, cultured at 37°C for 16 hours, took 100 μL of the enrichment solution after the enrichment culture, and diluted it appropriately and applied it to Salmonella, Staphylo...

Embodiment 3

[0027] Weigh 7.5g of bovine brain extract powder, 10g of bovine heart extract powder, 10g of peptone, 5g of sodium chloride, 2g of glucose, 2.5g of disodium hydrogen phosphate, 2g of mannitol, 2g of sodium pyruvate, 2g of glycine, 2g of escin, 1000ml of distilled water, stir to dissolve, adjust the pH value to 7.4 with 10 mol / L NaOH solution, autoclave at 121°C for 20min, and store at 4°C.

[0028] The cell concentration was 10 9 CFU / mL Salmonella, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Vibrio parahaemolyticus were diluted 10 7times, and then take 100 μL respectively, and add 10 mL of the prepared compound enrichment medium for Salmonella, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Vibrio parahaemolyticus to make the medium The concentration of each bacterium in the medium was 1 CFU / mL, cultured at 37°C for 16 hours, took 100 μL of the enrichment solution after the enrichment culture, and diluted it appropriately and appl...

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PUM

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Abstract

The invention discloses a composite enrichment medium for salmonella, staphylococcus aureus, escherichia coli, listeria monocytogenes and vibrio parahaemolyticus and a preparation method for the composite enrichment medium. The composite enrichment medium comprises the following components in parts by weight: 5-10 parts of ox brain extract powder, 5-15 parts of ox heart extract powder, 5-15 parts of peptones, 2.5-7.5 parts of sodium chloride, 1-3 parts of glucose, 1.5-3.5 parts of disodium hydrogen phosphate, 1-3 parts of mannitol, 1-3 parts of sodium pyruvate, 1-3 parts of glycine, 1-3 parts of aesculin, and 1000 parts of distilled water, and a pH value of the composite enrichment medium is 7.2-7.5. A preparation method for the composite enrichment medium comprises the following steps: adding the components into distilled water according to the formula, stirring and dissolving the components, regulating the pH value of the mixture, and sterilizing the mixture at high pressure. The five bacteria are main pathogenic bacteria which are needed to be detected according to the national food hygienic standard. The composite enrichment medium can be used for realizing rapid proliferation of the five pathogenic bacteria. After carrying out bacteria enrichment for 16 hours, the concentration of the thallus can be increased to 10<6>CFU / mL-10<9>CFU / mL from 1CFU / mL, so that the thallus can be directly used for carrying out high flux detection on multiple PCRs (polymerase chain reactions), gene chips and micro-fluidic chips, and thus, screening of the five pathogenic bacteria is completed.

Description

technical field [0001] The invention belongs to a pre-enrichment culture medium for pathogenic bacteria, in particular to a culture for compound enrichment of Salmonella, Staphylococcus aureus, Escherichia coli, Listeria monocytogenes and Vibrio parahaemolyticus at the same time Bases and their preparation methods. Background technique [0002] Foodborne pathogens are an important factor in the outbreak of food poisoning and foodborne diseases, and are an important risk and hidden danger of food safety. According to the report of the World Health Organization (WHO), there are as many as 150 million cases of diarrheal diseases worldwide every year, 70% of which are related to food contaminated by various pathogenic microorganisms. According to research data in my country, foodborne diseases caused by microbial pathogens account for 46.4%, seriously endangering people's health and causing major economic losses. Salmonella, Staphylococcus aureus, Escherichia coli, Listeria mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/42C12R1/445C12R1/19C12R1/63C12R1/01
Inventor 张岩刘铭李云葛少林邢婉丽
Owner CAPITALBIO CORP
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