Culture medium for composite enrichment of salmonella, Vibrio parahaemolyticus and Vibrio cholerae, and preparation thereof

A technology for Vibrio hemolyticus and Vibrio cholerae, which is applied in biochemical equipment and methods, determination/inspection of bacteria and microorganisms, etc., can solve the problem that the culture medium is not selective, the detection efficiency is low, and it does not conform to the development trend of pathogenic bacteria. and other problems to achieve the effect of shortening the enrichment time

Inactive Publication Date: 2009-04-22
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, for Salmonella, Vibrio parahaemolyticus and Vibrio cholerae, the combination of conventional bacterial enrichment culture, biochemical identification and automatic enzyme-linked fluorescent immunoassay method (VIDAS) is mainly used, and most of them take 4-6 days, and the procedures are complicated. Various, time-consuming and laborious, low detection efficiency, low detection sensitivity, and serious false negatives
In recent years, new technologies such as enzyme-linked fluorescent immunoassay (VIDAS), polymerase chain reaction (PCR), gold standard test paper method, API method, polymerase immunoassay (EIA) and DNA probe technology have been gradually applied to microorganisms. Detection greatly improves the sensitivity of detection and simplifies the operation. There are even detection methods for detecting multiple microorganisms on the same detection platform such as multiplex PCR technology, protein or antibody microarray receptors, immunoabsorption and DNA microarray technologies, but all these methods Still inseparable from selective enrichment and / or pre-enrichment
The existing enrichment method is mainly to carry out the respective enrichment treatment according to the strains to be monitored, that is, different strains use their own selective enrichment medium for the enrichment treatment, which is time-consuming and laborious, and does not conform to the modern detection method-platform simultaneous detection The development trend of multiple pathogenic bacteria
[0005] A universal enrichment medium (UPB) has been developed internationally that can proliferate a variety of bacteria at the same time. However, since this medium is not selective, in addition to the proliferation of the target bacteria to be monitored, other miscellaneous bacteria can also grow in large quantities. Miscellaneous bacteria A large number of growth will inevitably affect the growth of the target bacteria, the effect of UPB is unsatisfactory

Method used

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  • Culture medium for composite enrichment of salmonella, Vibrio parahaemolyticus and Vibrio cholerae, and preparation thereof
  • Culture medium for composite enrichment of salmonella, Vibrio parahaemolyticus and Vibrio cholerae, and preparation thereof
  • Culture medium for composite enrichment of salmonella, Vibrio parahaemolyticus and Vibrio cholerae, and preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Weigh 20.1g of buffered peptone water, 1.25g of glucose, 1.25g of mannitol, 0.05g of sodium pyruvate, 1.25g of anhydrous sodium sulfite, 5.0g of sodium citrate, 15.0g of sodium chloride, 2.5g of bile salt, and 1000ml of distilled water and heat Sterilize at 121°C for 18 minutes after melting. After the culture cools down to below 50°C, add 1.0 mg of potassium tellurite under sterile conditions, dissolve and mix well, and distribute for use.

[0021] Three target bacteria Salmonella, Vibrio parahaemolyticus and Vibrio cholerae and non-target bacteria Escherichia coli, Bacillus cereus, Yersinia enterica, Proteus, Enterobacter aerogenes, Shigella flexneri, Abalone Slant cultures of Shigella aeruginosa, Pseudomonas aeruginosa, Vibrio vulnificus, Vibrio alginolyticus will be diluted to 10 -4 Add 0.2ml of each target bacterium and non-target bacterium solution into the above-mentioned medium, and incubate at 37°C for 24h. The OD value at 540 nm was measured with a visible s...

Embodiment 2

[0027] Weigh 10.05g of buffered peptone water, 1g of glucose, 1g of mannitol, 0.03g of sodium pyruvate, 0.5g of anhydrous sodium sulfite, 1.0g of sodium citrate, 5.0g of sodium chloride, 1g of bile salt, and 1000ml of distilled water after heating and melting for 123 Sterilize at ℃ for 16 minutes. After the culture cools down to below 50°C, add 0.5 mg of potassium tellurite under aseptic conditions, dissolve and mix well, and distribute for use.

[0028] Three target bacteria Salmonella, Vibrio parahaemolyticus and Vibrio cholerae and non-target bacteria Escherichia coli, Bacillus cereus, Yersinia enterica, Proteus, Enterobacter aerogenes, Shigella flexneri, Abalone Slant cultures of Shigella aeruginosa, Pseudomonas aeruginosa, Vibrio vulnificus, Vibrio alginolyticus will be diluted to 10 -4 Add 0.2ml of each target bacterium and non-target bacterium solution into the above-mentioned medium, and incubate at 37°C for 24h. The OD value at 540 nm was measured with a visible spe...

Embodiment 3

[0034] Weigh 30.15g of buffered peptone water, 5.0g of glucose, 5.0g of mannitol, 0.05g of sodium pyruvate, 2.5g of anhydrous sodium sulfite, 5.0g of sodium citrate, 25.0g of sodium chloride, 5g of bile salt, and 1000ml of distilled water and heat to melt Then sterilize at 125°C for 15 minutes. After the culture cools down to below 50°C, add 2.5 mg of potassium tellurite under sterile conditions, dissolve and mix well, and distribute for use.

[0035] Diluted into three target bacteria Salmonella, Vibrio parahaemolyticus and Vibrio cholerae and non-target bacteria Escherichia coli, Bacillus cereus, Yersinia enterica, Proteus, Enterobacter aerogenes, Shigella flexneri bacteria, Shigella baumannii, Pseudomonas aeruginosa, Vibrio vulnificus, Vibrio alginolyticus slant culture, will be diluted to 10 -4 Add 0.2ml of each target bacterium and non-target bacterium solution into the above-mentioned medium, and incubate at 37°C for 24h. The OD value at 540 nm was measured with a visi...

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Abstract

The invention relates to a complex enrichment medium used for salmonella, vibrio parahaemolyticus and comma bacillus and a method for preparing the same. The formulation components of the complex enrichment medium in weight portion are as follows: 0.5 to 1.5 portions of buffer peptone water, 1000 portions of distilled water, 1.0 to 5.0 portions of glucose, 1.0 to 5.0 portions of mannitol, 0.03 to 0.06 portion sodium pyruvate, 0.5 to 2.5 portions of anhydrous sodium sulfite, 1.0 to 10.0 portions of sodium citrate, 5.0 to 25.0 portions of sodium chloride, 1 to 5 portions of cholate, and 0.0005 to 0.0025 portion of potassium tellurite. The method comprises the following steps: firstly, adding the buffer peptone water and the like into the distilled water, and sterilizing the mixture after heating and melting; and secondly, cooling the mixture to 50 DEG C, and then adding the potassium tellurite into the mixture to be mixed evenly. The complex enrichment medium can integrate two steps of primary enrichment and selective enrichment of bacterial detection to shorten the enrichment time to 24 hours, can perform enrichment on three target pathogens at the same time, and can inhibit the growth of other microorganisms.

Description

technical field [0001] The invention belongs to a pre-enrichment medium for pathogenic bacteria, in particular to a medium for compound enrichment of Salmonella, Vibrio parahaemolyticus and Vibrio cholerae at the same time. Background technique [0002] my country is a big exporter of aquatic products, but with the globalization of trade, my country's fishery will face unprecedented new problems and new challenges, and put forward higher requirements for the quality of my country's aquatic products. Pollution-free and pollution-free aquatic products will surely become the international market for aquatic products. Market entry ticket Contaminated pathogenic bacteria in food is one of the main factors causing food-borne diseases, and it is a huge and expanding worldwide public health problem. Controlling microbial contamination is currently the main content of solving the problem of aquatic product pollution . [0003] Salmonella, Vibrio parahaemolyticus and Vibrio cholerae ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/68C12Q1/04
CPCY02A50/30
Inventor 肖性龙余以刚吴晖李苏龙覃倚莹许喜林
Owner SOUTH CHINA UNIV OF TECH
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