HiMedia Laboratories GmbH, Marie-Curie-Str. 3, 64683, Einhausen, Germany.
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Intended Use:
Recommended for differentiation of Proteus and Providencia group of organisms from other members of Enterobacteriaceae on the basis of their ability to form phenyl pyruvic acid from phenylalanine.
Composition**
| Ingredients | g/L |
|---|---|
| Yeast extract | 3.000 |
| Sodium chloride | 5.000 |
| DL-Phenylalanine | 2.000 |
| Disodium hydrogen phosphate | 1.000 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.3±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Streak the test inoculum aseptically into the slant and incubate at appropriate conditions. Incubate the slants at 30-35°C for 18-24 hours.
Principle And Interpretation
The ability of Proteus species to convert phenylalanine to phenylpyruvic acid is an important reaction in the differentiation of Enterobacteriaceae (1,2). Based on this criterion, Buttiaux developed Phenylalanine Agar for differentiation of Proteus and Providencia group from other members of Enterobacteriaceae (3,4) by the ability of organism in the genera within Proteus to deaminate phenylalanine. Phenylalanine Agar is the modification of the medium originally developed by Ewing et al (5).
Yeast extract in the medium supports the growth of the organisms. Sodium chloride maintains osmotic equilibrium. The phenylalanine serves as the substrate for enzymes, which are able to deaminate it to form phenylpyruvic acid. A recommended technique is to inoculate the slant surface with plenty of inoculum and incubate it for 12-16 hours. After incubation, add 0.2 ml of 10% ferric chloride solution so that the solution floods all over the growth. The addition of (0.2 ml 3-5 drops) of a 10% aqueous ferric chloride solution (or a 12% aqueous ferric chloride solution acidified with 2.5 ml of concentrated HCl per 100 ml of reagent) to the cultures following incubation results in the appearance of a light to deep green color (positive reaction) or no color change (negative reaction). In a positive reaction, any phenylpyruvic acid present will react with the ferric salt in the reagent to give a green color. Interpret the results within 5 minutes upon addition of reagent as the green colour fades quickly (2,4).
Type of specimen
Isolated microorganism
Specimen Collection and Handling:
A recommended technique is to inoculate the slant surface with plenty of inoculum and incubate it for 12-16 hours. After incubation, add 0.2 ml of 10% ferric chloride solution so that the solution floods all over the growth. The addition of (0.2 ml 3-5 drops) of a 10% aqueous ferric chloride solution (or a 12% aqueous ferric chloride solution acidified with 2.5 ml of concentrated HCl per 100 ml of reagent) to the cultures following incubation results in the appearance of a light to deep green color (positive reaction) or no color change (negative reaction). In a positive reaction, any phenylpyruvic acid present will react with the ferric salt in the reagent to give a green color. Interpret the results within 5 minutes upon addition of reagent as the green colour fades quickly (2,4).
After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions:
In vitro diagnostic use only. For professional use only. Read the label before opening the container. Wear protective gloves/ protective clothing/eye protection/ face protection. Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established guidelines should be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations:
- Some organism may show poor growth due to nutritional variation.
- Other biochemical and serological testing must be performed for confirmation
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.
Quality Control
Appearance
Sterile Phenylalanine Agar Slant in glass tube.
Colour of medium
Yellow coloured slant
Quantity of medium
8ml of medium in glass tube
pH
7.10-7.50
Sterility Check
Passes release criteria
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours
| Organism | Inoculum (CFU) | Growth | Phenylalanine deaminase |
|---|---|---|---|
| # Klebsiella aerogenes ATCC 13048 (00175*) | 50-100 | luxuriant | negative reaction |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | luxuriant | negative reaction |
| Proteus mirabilis ATCC 25933 | 50-100 | luxuriant | positive reaction, green colouration after addition of 10% ferric chloride |
| Proteus vulgaris ATCC 13315 | 50-100 | luxuriant | positive reaction, green colouration after addition of 10% ferric chloride |
| Providencia alcalifaciens ATCC 9886 | 50-100 | luxuriant | positive reaction, green colouration after addition of 10% ferric chloride |
Key: *Corresponding WDCM numbers.
(#) Formerly known as Enterobacter aerogenes
Storage and Shelf Life
Store between 2-8°C. Use before expiry date on the label. Product performance is best if used within stated expiry period.
Disposal
User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with sample must be decontaminated and disposed of in accordance with current laboratory techniques (6,7).
| Product Name | Phenyl Alanine Agar Slant |
|---|---|
| SKU | SL044 |
| Customized Product Available | No |
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