PC (PSEUDOMONAS CEPACIA) AGAR

Cat. no. G48 PC Agar, 15x100mm Plate, 18ml 10 plates/bag

INTENDED USE

Hardy Diagnostics PC ( Pseudomonas cepacia ) Agar is recommended for the selective isolation of Burkholderia ( Pseudomonas ) cepacia from clinical and environmental materials.

SUMMARY

PC Agar was developed by Gilligan, et al. (1) They were able to demonstrate that P. cepacia could be recovered from mixed cultures of cystic fibrosis patients using this media. This formula contains bile salts and crystal violet, which inhibits gram-positive bacteria. Gram-negative bacteria can be inhibited by the addition of polymyxin B and ticarcillin. P. cepacia utilizes the pyruvate and produces alkaline products that turns the media bright pink to red.

FORMULA

Ingredients per liter of deionized water:*

Sodium Pyruvate 5.0gm
Dipotassium Phosphate 4.3gm
Monopotassium Phosphate 2.1gm
Peptic Digest of Animal Tissue 1.0gm
Ammonium Sulfate 1.0gm
Bile Salts No. 3 0.5gm
Magnesium Sulfate 0.2gm
Ticarcillin 0.1gm
Phenol Red 20.0mg
Ferrous Ammonium Sulfate 10.0mg
Crystal Violet 1.0mg
Polymyxin B 300,000U
Agar 15.0gm

Final pH 7.1 +/- 0.3 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-8ºC. Media should not be used if there are any signs of deterioration (shrinking, cracking, or discoloration), contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.

PRECAUTIONS

PROCEDURE

Specimen Collection: Consult listed references for information on specimen collection. (1-3,5,6) Infectious material should be submitted directly to the laboratory without delay and protected from excessive heat and cold. If there is to be a delay in processing, the use of a buffered holding medium has been shown effective in the recovery of most microorganisms.

Method of Use: Allow the plates to warm to room temperature, and the agar surface to dry before inoculating. Inoculate and streak the specimen as soon as possible after collection. If the specimen to be cultured is on a swab, roll the swab over a small area of the agar surface. Streak for isolation with a sterile loop. Incubate plates aerobically at 35ºC. for 18-72 hours. Examine plates daily for typical colonial morphology and growth characteristics.

INTERPRETATION OF RESULTS

Growth of gray-white colonies surrounded by bright pink to red zone in medium may be presumptively identified as Burkholderia ( Pseudomonas ) cepacia.

LIMITATIONS

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, other culture media, swabs, applicator sticks, incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Burkholderia (Pseudomonas) cepacia
ATCC ® 25416
A 18-72hr 35°C Aerobic Growth; gray-white colonies surrounded by a bright pink to red zone in medium
Pseudomonas aeruginosa
ATCC ® 27853
B 24hr 35°C Aerobic Inhibited
(partial to complete)
Escherichia coli
ATCC ® 25922
B 24hr 35°C Aerobic Inhibited
(partial to complete)
Staphylococcus aureus
ATCC ® 25923
B 24hr 35°C Aerobic Inhibited
(partial to complete)

User Quality Control

Physical Appearance

PC Agar should appear clear to slightly hazy, and orange-red in color.

B. cepacia growing on PC Agar

Burkholderia (Pseudomonas) cepacia (ATCC ® 25416) colonies growing on PC Agar (Cat. no. G48). Incubated aerobically for 48 hours at 35ºC.

B. cepacia growing on PC Agar

Uninoculated plate of PC Agar (Cat. no. G48).

REFERENCES

1. Gilligan, P.H., et al. 1985. J. Clin. Microbiol.; 22:5.

2. Anderson, N.L., et al. Cumitech 3B; Quality Systems in the Clinical Microbiology Laboratory, Coordinating ed., A.S. Weissfeld. American Society for Microbiology, Washington, D.C.

3. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

4. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

5. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

6. Koneman, E.W., et al. Color Atlas and Textbook of Diagnostic Microbiology, J.B. Lippincott Company, Philadelphia, PA.


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