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Selective isolation medium for enterobacter sakazakii

A technology of Enterobacter sakazakii and culture medium, which is applied in the field of selective isolation medium of Enterobacter sakazakii, can solve the problems of low work efficiency, unfavorable separation of suspicious colonies, long detection time, etc., and achieve the effect of improving work efficiency

Inactive Publication Date: 2008-05-28
SHENZHEN ACAD OF METROLOGY & QUALITY INSPECTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome that the existing culture medium (such as VRBGA) has no selectivity or poor selectivity to Enterobacter sakazakii; Enterobacter sakazakii spreads and grows therein into mucus and is not conducive to the separation of suspicious colonies; the detection time is long and the work efficiency is low. Inferior disadvantages

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The formula of the Enterobacter sakazakii culture medium of Example 1 is shown in Table 1. Mix 33.0g of brain heart powder, 4.0g of sodium chloride, 1.0g of agar and 1000mL of water, heat to dissolve, distribute them in appropriate containers, autoclave at 121°C for 3min to obtain the basic medium, and store it in a sealed container for later use . Accurately weigh 3.0 mg of vancomycin, mix it with 1 mL of sterile water, and filter and sterilize the stock solution of vancomycin. Accurately weigh 0.08 mg of cefalotin, mix it with 1 mL of phosphate buffer (pH 6.0, 0.1 mol / L), and filter and sterilize to obtain a cefalotin storage solution.

[0023] Immediately before use, heat to dissolve the basal medium, then cool to 60°C, add 1mL vancomycin stock solution, 1mL cephalothin stock solution and 0.50g 5-bromo-4chloro-3-indole-aD glucoside in turn, shake Evenly, pour a 9cm diameter plate, 10-15mL / plate. Separately add 10 μl of Staphylococcus aureus, Listeria monocytogenes, Shige...

Embodiment 2

[0026] Repeat the method of Example 1, except that the composition of the basic medium, the composition of the vancomycin storage solution, the composition of the cefalotin storage solution and the added 5-bromo-4chloro-3-indole-α-D glucoside are different, such as As shown in Table 1, the basic medium is composed of 38.0g brain heart extract powder, 5.0g sodium chloride, 1.5g agar and 1000mL water; the vancomycin storage solution is made up of 4.0mg vancomycin and 1mL sterile water; The cefalotin storage solution is made up of 0.10mg cefalotin and 1mL sterile water; the added 5-bromo-4chloro-3-indole-α-D glucoside is 0.10g.

[0027] After 24 hours of culture at 37°C, normal pressure and air, the growth of the colony was observed by naked eyes. The results are shown in Table 2.

[0028] Table 2 shows that, except for Enterobacter aerogenes, the other 9 non-sakazakii Enterobacter strains did not grow. Although Enterobacter aerogenes grows, its colony is earthy yellow, and it is eas...

Embodiment 3

[0030] Repeat the method of Example 1, except that the composition of the basic medium, the composition of the vancomycin storage solution, the composition of the cefalotin storage solution and the added 5-bromo-4chloro-3-indole-α-D glucoside are different, such as As shown in Table 1, the basal medium is composed of 43.0g brain heart extract powder, 6.0g sodium chloride, 2.0g agar and 1000mL water; the vancomycin storage solution is made up of 5.0mg vancomycin and 1mL sterile water; Cefalotin storage solution is made up of 0.15mg cefalotin and 1mL sterile water; the added 5-bromo-4chloro-3-indole-α-D glucoside is 0.08g.

[0031] After 24 hours of culture at 37°C, normal pressure and air, the growth of the colony was observed by naked eyes. The results are shown in Table 2.

[0032] Table 2 shows that, except for Enterobacter aerogenes, the other 9 non-sakazakii Enterobacter strains did not grow. Although Enterobacter aerogenes grows, its colony is earthy yellow, and it is easy to...

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Abstract

Provided is a selective separation growth medium of enterobacter sakazakii. The growth medium is composed of a basic growth medium, antibiotics and color-developing agent, wherein the basic growth medium is composed of the following components of 33.0-43.0 grams of brain heart impregnating powder, 4.0-6.0 grams of sodium chloride, 1.0-2.0 grams of agar and 1000 milliliters of water, the antibiotics is vanoomycin, cefalotin, or the combination of vanoomycin and cefalotin, and the color-developing agent is 5-bromine-4-chlorine 3-indole-alpha-D glucoside. The growth medium of the enterobacter sakazakii has the advantages that the selectivity is high, the hypertrophy of the enterobacter sakazakii is inhibited, the detection time is shortened, the operating efficiency is increased and the like, which are relative to VRBGA growth medium.

Description

Technical field [0001] The present invention relates to a medium, and more specifically, to a medium for selective separation of Enterobacter sakazakii. Background technique [0002] Enterobacter sakazakii is a gram-negative non-bacillus bacillus with pericytes and can move. Infants and young children are often infected by eating milk powder contaminated by Enterobacter sakazakii. The clinical manifestations are meningitis, sepsis, necrotizing colitis and other symptoms, and the mortality rate is high. [0003] The current medium used to isolate Enterobacter sakazakii is the industry standard SN / T1632.1-2005 or VRBGA recommended by the US FDA. Enterobacter sakazakii can grow into two colonies on the VRB GA plate, one is Yiwei The typical smooth colony moving in the inoculation ring, the other is a dry or slimy colony, but most of it grows into a slimy red colony with bile acid deposits around it. At the same time, some other Enterobacteriaceae can also grow into colonies similar ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20
Inventor 李翔杨万颖朱崇全罗美中杨国武李传礼祝仁发赖心田
Owner SHENZHEN ACAD OF METROLOGY & QUALITY INSPECTION
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