COLUMBIA BLOOD AGAR
|Cat. no. A16||Columbia Blood Agar, 15x100mm Plate, 17ml||10 plates/bag|
|Cat. no. A16BX||Columbia Blood Agar, 15x100mm Plate, 17ml||100 plates/box|
Hardy Diagnostics Columbia Blood Agar is recommended for use as a general growth medium for the cultivation and hemolytic determination of fastidious and non-fastidious microorganisms from clinical specimens.
Columbia Blood Agar was first described in 1966 by Ellner, Stoessel, Drakeford, and Vasi who incorporated animal derived peptone, enzymatic digests of casein, and defibrinated sheep blood into one medium. (3) It was found to be an improved blood agar, promoting both luxuriant and rapid growth, improved pigment production, typical colony morphology, and sharply defined hemolytic reactions.
Ingredients per liter of deionized water:*
|Enzymatic Digests of Casein||10.0gm|
|Casein Yeast Peptone||10.0gm|
|Tryptic Digest of Beef Heart||3.0gm|
Final pH 7.3 +/- 0.2 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.
Specimen Collection: Consult listed references for information on specimen collection. (1,2,4,6) 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 specimen should be inoculated onto an appropriate transport media and refrigerated until inoculation.
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 or in a 5% to 10% carbon dioxide atmosphere at 35-37ºC. for 18-24 hours. Examine colonial morphology, characteristics, and hemolytic reactions.
INTERPRETATION OF RESULTS
Consult listed references for the identification of colony morphology and further biochemical tests required for identification. (1,2,4,6)
Some beta-hemolytic streptococci strains will develop green hemolytic zones on Columbia Blood Agar. It is recommended to subculture all such streptococci strains onto Blood Agar plates (Cat. no. A10) to verify this reaction.
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.
|Test Organisms||Inoculation Method*||Incubation||Results|
ATCC ® 19615
|A||24hr||35°C||Aerobic 2 **||Growth, beta-hemolysis|
ATCC ® 6305
|A||24hr||35°C||Aerobic 2 **||Growth, alpha-hemolysis|
ATCC ® 25923
|A||24hr||35°C||Aerobic 2 **||Growth|
ATCC ® 25922
|A||24hr||35°C||Aerobic 2 **||Growth|
User Quality Control
Columbia Blood Agar should appear opaque, and cherry red in color.
Streptococcus pyogenes (ATCC ® 19615) colonies growing on Columbia Blood Agar (Cat. no. A16). Incubated aerobically for 24 hours at 35ºC.
Streptococcus pneumoniae (ATCC ® 6305) colonies growing on Columbia Blood Agar (Cat. no. A16). Incubated aerobically for 24 hours at 35ºC.
Staphylococcus aureus (ATCC ® 25923) colonies growing on Columbia Blood Agar (Cat. no. A16). Incubated aerobically for 24 hours at 35ºC.
Escherichia coli (ATCC ® 25922) colonies growing on Columbia Blood Agar (Cat. no. A16). Incubated aerobically for 24 hours at 35ºC.
Uninoculated plate of Columbia Blood Agar (Cat. no. A16).
1. 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.
5. Tille, P.M., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.
3. Ellner, et al. 1966. Am. Journ. Clin. Path. ; 45:502.
4. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.
5. MacFaddin, J.F. 1985. Media for Isolation, Cultivation, Identification, Maintenance of Bacteria , Vol. I. Williams & Wilkins, Baltimore, MD.
6. Versalovic, J., et al. Manual of Clinical Microbiology. American Society for Microbiology, Washington, D.C.
7. Quality Assurance for Commercially Prepared Microbiological Culture Media , M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.
ATCC is a registered trademark of the American Type Culture Collection.