Coloring culture medium for detecting food-borne pathogenic Yersinia

A technology of Yersinia and chromogenic culture medium, which is applied in the field of food safety microbial inspection and monitoring, can solve the problems of high cost, unfavorable promotion and application, and high price, and achieve simple result judgment, wide application prospects, and easy operation Effect

Active Publication Date: 2017-04-26
BEIJING JUNLIKANG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] For the inspection method of Yersinia enterocolitica in food, the traditional method is commonly used at present. Taking the method of food safety standard GB4789.8 as an example, the specific operation is as follows: after the pre-enrichment and the treatment of the enrichment solution, inoculate CIN-1 and improved Y medium were used for culture and separation, and then the suspicious bacteria were tested for Klebsiella iron, urease, semi-solid dynamic test and microscopic examination to obtain the results of whether they belong to the genus Yersinia. It takes at least 6-7 days for biochemical reactions to distinguish Yersinia enterocolitica, and the efficiency is low
In addition, the detection of Yersinia enterocolitica and other Yersinia of this genus also uses PCR technology and other methods, which require special equipment and professional technicians, and the cost is high, which is not conducive to popularization and application. Currently, there is no food source Specific method for the sexual pathogen Yersinia pseudotuberculosis
[0005] The bacterial biochemical identification technology based on the traditional culture medium by adding specific enzyme substrates can be better connected with the traditional method at present, and the cultivation, separation and identification can be completed at one time, the operation is simple, the detection time is greatly shortened, and through The characteristic identification of target bacteria improves the accuracy, so it has broad application prospects. It has been applied to bacteria such as Salmonella, Vibrio, Listeria, and pathogenic E. coli O157 in many fields such as clinical medicine, food hygiene, and environmental protection. For the detection of Yersinia enterocolitica, CHROMagar has developed a chromogenic medium (CHROMagarY. Yersinia is the target, that is, Yersinia enterocolitica is divided into 6 groups according to phenotypic characteristics, and the chromogenic medium developed by CHROMagar can detect the remaining 5 groups with strong pathogenicity except 1A Enterocolitica Yersinia (groups 1B, 2, 3, 4, 5), that is, the target bacteria that can be isolated and identified by this medium are not all Yersinia enterocolitica, and cannot be isolated and detection of Yersinia pseudotuberculosis
The purpose of food safety inspection is to reduce the risk. As far as Yersinia enterocolitica is concerned, taking the method of food safety standard GB4789.8 as an example, the clear scope of inspection is all biological groups. Therefore, this product cannot meet the requirements of food The demand for safety testing, in addition, in the application of medical testing, its expensive price increases the cost of the user department

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  • Coloring culture medium for detecting food-borne pathogenic Yersinia
  • Coloring culture medium for detecting food-borne pathogenic Yersinia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: Specificity experiment of Yersinia chromogenic medium of the present invention to Yersinia

[0032] 1) Solid plate preparation:

[0033] According to the formula, each 1000mL medium contains 15g soybean peptone, 5g yeast powder, 5g sodium chloride, 2g sodium oxalate, 2g No. 3 bile salt, 13g agar, 5 bromo-4-chloro-3-indole-β- 0.05 g of D-ribofuranoside, 0.05 g of 5-bromo-6-chloro-3-indole-β-D-galactopyranoside, the balance is water, and the pH is 7.4±0.2 to prepare a culture medium. Add the components of the above-mentioned chromogenic medium to deionized water, stir to dissolve, adjust the pH to 7.4±0.2, heat to boil, cool to 45~55°C, perform aseptic operation, pour into a plate, and set aside;

[0034] 2) Activation and cultivation of bacteria:

[0035]The tested bacteria included Staphylococcus aureus ATCC25923, enterotoxigenic Escherichia coli IQCC10113, Enterobacter sakazakii ATCC29544, Salmonella typhimurium ATCC14028, Aeromonas veterinus ATCC51108,...

Embodiment 2

[0045] Embodiment 2: Selective effect experiment of Yersinia chromogenic medium of the present invention

[0046] 1) Solid plate preparation:

[0047] According to the formula, each 1000mL medium contains 15g soybean peptone, 5g yeast powder, 5g sodium chloride, 2g sodium oxalate, 2g No. 3 bile salt, 13g agar, 5 bromo-4-chloro-3-indole-β- D-ribofuranoside 0.05g, 5-bromo-6-chloro-3-indole-β-D-galactopyranoside 0.05g, cephalosporin 15mg, novobiocin 2.5mg, the balance is water, the pH is 7.4 ± 0.2 Preparation of medium. Add the components of the above-mentioned chromogenic medium except antibiotics into deionized water, stir to dissolve, adjust the pH to 7.4±0.2, heat to boil, wait to cool to 45~55°C, add corresponding antibiotics, perform aseptic operation, pour tablet, spare;

[0048] 2) Activation and cultivation of bacteria:

[0049] See Example 1 for the standard strains tested.

[0050] The above-mentioned strains were respectively activated and inoculated on Columbia ...

Embodiment 3

[0058] Embodiment 3: the Yersinia chromogenic culture medium of the present invention distinguishes strongly pathogenic Yersinia enterocolitica

[0059] 1) Solid plate preparation:

[0060] According to the formula, each 1000mL medium contains 15g soybean peptone, 5g yeast powder, 5g sodium chloride, 2g sodium oxalate, 2g No. 3 bile salt, 13g agar, 0.5g aescin, 0.5g ferric ammonium citrate, 5 Bromo-4-chloro-3-indole-β-D-ribofuranoside 0.05g, 5-bromo-6-chloro-3-indole-β-D-galactopyranoside 0.05g, cephalosporin 15mg, Novobiocin 2.5mg, the balance is water, and the pH is 7.4±0.2 to prepare the culture medium. Add the components of the above chromogenic medium except antibiotics into deionized water, stir to dissolve, adjust the pH to 7.4±0.2, heat to boil, wait to cool to 45~55°C, add the corresponding antibiotics, perform aseptic operation, pour tablet, spare;

[0061] 2) Activation and cultivation of bacteria:

[0062] The tested bacteria include Staphylococcus aureus ATCC2...

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Abstract

The present invention discloses a coloring culture medium for detecting food-borne pathogenic Yersinia, wherein the culture medium contains soybean peptone, yeast powder, sodium chloride, sodium oxalate, agar, a beta-riboside chromogenic substrate, a beta-galactoside chromogenic substrate, esculin, ammonium ferric citrate, bile salt No.3, cephalosporin, novobiocin, and the balance of water. According to the present invention, the coloring culture medium has characteristics of simple formula and convenient preparation operation, can be used for the separation and the detection of food-borne pathogenic Yersinia, and further has advantages of specificity, high efficiency, rapidness, low cost, simple result determination, wide application prospect, and the like.

Description

technical field [0001] The invention relates to a culture medium for microorganism detection, in particular to a chromogenic culture medium for detecting food-borne pathogenic Yersinia, belonging to the field of food safety microorganism detection and monitoring. Background technique [0002] Yersinia enterocolitica is one of the important food-borne pathogenic bacteria, which can survive and proliferate in low-temperature preserved food, and can cause extremely serious food poisoning incidents. In some countries, the degree even exceeds Shigella, close to Salmonella and Campylobacter jejuni. Once infected, it will cause a variety of diseases and seriously threaten people's health. It is a routine inspection item for food safety in various countries. my country's food safety standard GB4789 .8 specifies the inspection method for Yersinia enterocolitica in food. [0003] Yersinia enterocolitica belongs to the genus Yersinia in taxonomy, and there are 11 species in this genus. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12R1/01
CPCC12Q1/045
Inventor 赵贵明陈颖姬庆龙王娉杨海荣
Owner BEIJING JUNLIKANG BIOTECHNOLOGY CO LTD
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