A method for isolating Staphylococcus aureus that eliminates the interference of food background bacteria

A separation method, the technology of staphylococcus, applied in the field of staphylococcus aureus separation, can solve the problems of hemolysis, background bacteria contamination, misjudgment, etc., and achieve the effect of improving the separation rate and eliminating the interference of bacteria

Active Publication Date: 2022-05-31
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when Staphylococcus aureus was isolated from frozen meat and meat products (especially frozen chicken) in the early stage, it was found that due to the presence of diffuse background bacteria contamination, the selective plate was often covered with a layer of gray matter, which made the appearance of the colony The morphology changes, which causes misjudgment and leads to a large number of false negative results
At the same time, the miscellaneous bacteria also appeared hemolysis phenomenon on the blood plate, which will produce a large number of false positive results
More importantly, due to the proliferation of miscellaneous bacteria, it is difficult to separate them from Staphylococcus aureus in subsequent verification experiments

Method used

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  • A method for isolating Staphylococcus aureus that eliminates the interference of food background bacteria
  • A method for isolating Staphylococcus aureus that eliminates the interference of food background bacteria
  • A method for isolating Staphylococcus aureus that eliminates the interference of food background bacteria

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Embodiment 1

[0029] A method for isolating Staphylococcus aureus that can eliminate the interference of background bacteria, comprising the following steps:

[0030] Step 1, strain activation and preparation of working bacterial solution:

[0031] Streak the strains stored in the glycerol tubes in the -80°C ultra-low temperature refrigerator to activate them on ordinary trypsin soybean agar plates, culture them statically at 37°C for 18-24 hours, and then store them in the refrigerator at 4°C. Pick a single colony and inoculate it into TSB, and place it in a shaker incubator at 37°C and 200 rpm for overnight culture. After the culture, centrifuge at 10000rpm for 30s, discard the supernatant, wash the precipitate with saline containing 0.85% sodium chloride, repeat twice, and finally adjust the OD with a microplate reader 600 Prepared as a working bacterial solution.

[0032] Step 2, determine the inoculation ratio:

[0033] Adjust ATCC 29213 and miscellaneous bacteria to a consistent OD...

Embodiment 2

[0055] The application of the Staphylococcus aureus enrichment and isolation method that can eliminate background miscellaneous bacteria interference includes the following steps:

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Abstract

The first aspect of the present invention discloses a method for isolating Staphylococcus aureus that eliminates the interference of food background bacteria. The separation method includes an enrichment method and a purification method. The enrichment method uses improved enrichment broth, and the purification method uses improved trypsin soybean agar medium; the second aspect of the present invention also discloses the application of a Staphylococcus aureus enrichment method and purification method in the isolation of Staphylococcus aureus; the third aspect of the present invention also discloses a method for isolating Staphylococcus aureus A Staphylococcus aureus isolation kit prepared by application. The method for isolating Staphylococcus aureus that eliminates the interference of food background bacteria effectively eliminates the interference of bacteria, improves the separation rate of Staphylococcus aureus in food, and has the advantages of low detection method cost, simple operation, high accuracy, etc. Advantages, broad application prospects and economic benefits.

Description

technical field [0001] The invention relates to the technical field of food safety detection, in particular to a method for isolating Staphylococcus aureus for eliminating the interference of food background bacteria. Background technique [0002] Staphylococcus aureus (Staphylococcus aureus) is an important food-borne pathogen, which often causes food poisoning, hospital and community-acquired infections, and is the key object of food-borne disease surveillance in my country. Because Staphylococcus aureus exists widely in nature and is zoonotic, it generally has a strong ability to form a bacterial film. Therefore, Staphylococcus aureus is considered to have the potential to spread drug resistance, and multi-drug resistance occurs from time to time. In recent years, as the problem of bacterial drug resistance has become increasingly serious, researchers have realized that multidrug-resistant Staphylococcus aureus contaminated in food may become a carrier of drug resistance ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/02C12R1/445
CPCC12N1/20C12N1/02Y02A40/90
Inventor 施春雷区楚君尚岱麒常江杨婧娴陈博
Owner SHANGHAI JIAOTONG UNIV
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