BURKHOLDERIA CEPACIA SELECTIVE AGAR (BCSA)

Cat. no. G09 Burkholderia cepacia Selective Agar (BCSA),
15x100mm Plate, 18ml
10 plates/bag

INTENDED USE

Hardy Diagnostics Burkholderia cepacia Selective Agar (BCSA) is used for the selective isolation of Burkholderia (Pseudomonas) cepacia .

SUMMARY

Burkholderia cepacia Selective Agar (BCSA) was developed by Henry, Campbell, LiPuma and Speert for the selective isolation of Burkholderia (Pseudomonas) cepacia .(2) Burkholderia cepacia is commonly isolated from cystic fibrosis patients. Correct identification of the organism is critical to patient care. (2) It was found that BCSA had a lower false-positivity rate than either Oxidation-Fermentation-Polymyxin-Bacitracin-Lactose (OFBPL) Agar or PC (Pseudomonas cepacia) Agar. This finding was confirmed later by Henry, Campbell, McGimpsey, Clarke, Louden, Burns, Roe, Vandamme and Speert.(1)

BCSA contains peptones and sugars that supply nutrients for the growth of Burkholderia cepacia, and other microorganisms. Crystal violet is added to inhibit growth of gram-positive organisms. Antimicrobics are incorporated to inhibit organisms other than Burkholderia cepacia.

FORMULA*

Agar 17.0gm
Sucrose 12.0gm
Lactose 12.0gm
Pancreatic Digest of Casein 13.0gm
Gentamicin 13.0gm
Yeast Extract 1.5gm
Phenol Red 0.08gm
Vancomycin 2.5mg
Crystal Violet 2.0mg
Sodium Chloride 1.0mg

Final pH 6.8 +/- 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. away from direct light. Media should not be used if there are any signs of deterioration (shrinking, cracking, or discoloration), hemolysis, 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. (1-4) 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 specimen should be inoculated onto an appropriate transport media and refrigerated until inoculation.

Method of Use: Allow plates to warm to room temperature. The agar surface should be 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 35-37ºC. for 24-72 hours. Protect from light. Examine plates for colonial morphology.

INTERPRETATION OF RESULTS

Following incubation, BCSA is examined for growth. Burkholderia cepacia colonies are typically translucent and rough. On BCSA, the growth of B. cepacia will cause a color change in the media from red-orange to yellow. Consult the listed references for further procedures for identification of isolates.(3-6)

LIMITATIONS

Organisms other than Burkholderia cepacia may grow on BCSA. (1) Colonies that grow may require further biochemical testing for complete identification.

Some strains of Burkholderia cepacia isolated from patients who have been colonized for a number of years may show minor changes in biochemical reactions. These strains may show good growth but may not change the color of the media to yellow. Further biochemical testing should be performed on isolates showing good growth but no color change.

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 24-72hr 35°C Aerobic Growth; color change in media from red-orange to yellow
Pseudomonas aeruginosa
ATCC® 27853
B 24hr 35°C Aerobic Partial to complete inhibition
Staphylococcus aureus
ATCC® 25923
B 24hr 35°C Aerobic Partial to complete inhibition

User Quality Control

PHYSICAL APPEARANCE

Burkholderia cepacia Selective Agar (BCSA) should appear slightly opalescent, and reddish-orange in color.

B. cepacia growing on BCSA

Burkholderia (Pseudomonas) cepacia (ATCC® 25416) colonies growing on Burkholderia cepacia Selective Agar (Cat. no. G09). Incubated aerobically for 24 hours at 35ºC.

P. aeruginosa inhibited on BCSA

Pseudomonas aeruginosa (ATCC® 27853) growth inhibited on Burkholderia cepacia Selective Agar (Cat. no. G09). Incubated aerobically for 24 hours at 35ºC.

REFERENCES

1. Henry, D., et. al. 1999. Comparison of Isolation Media for Recovery of Burkholderia cepaciaComplex from Respiratory Secretions of Patients with Cystic Fibrosis. J. Clin. Micro.; 37:1004-1007.

2. Henry, D., et. al. 1997. Identification of Burkholderia cepacia Isolates from Patients with Cystic Fibrosis and Use of a Simple New Selective Medium. J. Clin. Micro.; 35:614-619.

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. 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.

7. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22-A2. Clinical Laboratory Standards Institute (CLSI - formerly NCCLS), Villanova, PA.


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121415hh