E. coli chromogenic medium
E. coli chromogenic medium is a modified medium from Qingdao Haibo Biotech Co., Ltd. for rapid detection of E. coli in food, water, milk, ice cream and meat products. Escherichia coli is blue-green, coliform is colorless, other bacteria are yellow or colorless, and Gram-positive bacteria are inhibited. Ingredients (g/L) Special nutrients This formula can be modified or added with nutrients to get good results. Note that this medium is for laboratory use only. references
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37.9
Reagent
0.2
Agar
12.0
pH 6.8 ± 0.2 25 °C
Usage Weigh 50.1g of this product into 1000ml distilled water, heat and dissolve without stirring, do not boil for more than 1 minute. Dispense the flask and autoclave at 121 °C for 15 minutes. Take 1ml of the prepared sample solution into the center of a sterile plate with a diameter of 9cm, pour into a medium of about 15ml cooled to 45 -50 °C, mix and solidify, and incubate at 36 ± 1 °C for 18 ~ 24h. Or when cooling to 45-50 °C, pour into a sterile plate. After the agar is completely solidified, place it in an incubator at 37 °C for 1-2 hours, add 1 ml of the appropriate dilution of the sample solution, and apply it to the medium. Incubate at 18 ± 1 °C for 18 to 24 hours.
Store prepared plates should be used immediately, and direct exposure to light should be avoided. The dry medium should be placed in a dark and dry place, and the storage temperature should be 2-8 °C.
Failure-dried media cannot be used beyond shelf life, agglomeration, and color change.
Steps
1. Prepare sample liquid according to national standards, SN standards, FDA standards or other methods.
2, take 1ml of the sample solution, add to the chromogenic medium cooled to 45-50 °C, mix or apply on the plate
3, 36 ± 1 °C culture for 18 ~ 24h, select a plate with 30 ~ 200 colonies, count the typical E. coli colonies appear on the plate. Typical colonies of E. coli are blue-green, and other bacteria are colorless or yellow colonies.
Total E. coli = blue-green colony
4. Suspicious E. coli colonies can be streaked onto nutrient agar plates, cultured at 36 ± 1 °C for 12-16 hours, and single colonies are picked for complete biochemical test of Escherichia coli (the company has 20 biochemical identification tube sets).
QC
1. Appearance The dry medium has a uniform powder with good fluidity and is milky white. The prepared plate was a clear, colorless solid medium.
2, microbial test at 36 ± 1 °C for 18 ~ 24h hours: quality control strain
ATCC
Growth condition colony color
Listeria monocytogenes
27853
-/+
colorless
Staphylococcus aureus
25923
-/+
colorless
E.coli
25922
+++
blue
salmonella
+++
colorless
Coliform
+++
colorless
Limitations of the method The chromogenic medium is used to identify E. coli. In a few cases, false positives may occur, but false negatives may not occur, and other supplemental tests may be required.
Typical characteristics E. coli typical colonies are blue-green, coliforms are colorless colonies, and other bacteria are colorless or yellow colonies.
FRAMPTON, EW, RESTAINO, L. a. BLASZKO, L.: Evaluation of ?-glucuronidase substrate in a 24 hour direct plating method for Escherichia coli. – J. Food Protection, 51; 402-404 (1988).
KILIAN, M. a. BüLOW, P.: Rapid diagnosis of Enterobacteriaceae. I. Detection of bacterial glycosidases. – Acta Pathol. Microbiol. Scand. Sect. B 84; 245-251 (1976).
LE MINOR, L. a. HAMIDA, F. BEN: Advantages de la recherche de la ?–galactosidase sur celle de la fermentation du lactose en milieu complexe dans le diagnostic bactériologique, en particulier des Enterobacteriaceae. – Ann. Inst. Pasteur ( Paris ), 102; 267-277 (1962).