COLUMBIA CNA AGAR

Cat. no. A50 Columbia CNA Agar, 15x100mm Plate, 19ml 10 plates/bag
Cat. no. A50BX Columbia CNA Agar, 15x100mm Plate, 19ml 100 plates/box
Cat. no. A129 Columbia CNA Agar, 86x128mm Omniplate, 30ml 10 plates/bag
Cat. no. GA50 Columbia CNA Agar, 15x100mm Plate, 19ml
(reduced stacking ring)
10 plates/bag
Cat. no. J52 Columbia CNA / EMB Agar, Levine, 15x100mm Biplate,
10ml/10ml
10 plates/bag
Cat. no. J62 Columbia CNA / MacConkey Agar, 15x100mm Biplate,
10ml/10ml
10 plates/bag
Cat. no. J66 Bile Esculin Agar (BEA) with Azide / Columbia CNA Agar,
15x100mm Biplate, 10ml/10ml
10 plates/bag
Cat. no. J116 Columbia CNA / BluEcoli™, 15x100mm Biplate, 10ml/10ml 10 plates/bag

INTENDED USE

Hardy Diagnostics Columbia CNA Agar is recommended for use as a selective growth medium for the isolation and differentiation of gram-positive cocci from clinical and non-clinical specimens which contain mixed flora.

Cat. no. A129 is not intended to be used for the diagnosis of human disease.

SUMMARY

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 form of Blood Agar, promoting both luxuriant and rapid growth, improved pigment production, typical colony morphology, and sharply defined hemolytic reactions. Ellner, et al., also described the use of nalidixic acid and colistin in Columbia Blood Agar. (3) Columbia CNA Agar was designed to suppress the growth of most gram-negative bacteria, including Klebsiella , Proteus , and Pseudomonas species from mixed flora specimens, thus isolating for gram-positive staphylococci and streptococci. (3)

FORMULA

Ingredients per liter of deionized water:*

Pancreatic Digests of Casein 12.0gm
Peptic Digest of Animal Tissue 5.0gm
Sodium Chloride 5.0gm
Yeast Extract 3.0gm
Beef Extract 3.0gm
Corn Starch 1.0gm
Colistin Sulfate 10.0mg
Nalidixic Acid 5.0mg
Sheep Blood 50.0ml
Agar 13.5gm

Final pH 7.3 +/- 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

For Cat. nos. A50, A50BX, GA50, J52, J62, J66, J116.

For Cat. no. A129.

PROCEDURE

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, anaerobically, or under conditions of increased CO 2 (5-10%) at 35-37ºC. for 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)

LIMITATIONS

Some beta-hemolytic streptococci strains will develop green hemolytic zones on Columbia CNA Agar. It is recommended to subculture all such streptococci strains onto Blood Agar plates (Cat. no. A10) to verify this reaction.

For optimal performance, it is recommended to incubate plates under increased CO 2 or aerobic conditions. Atmosphere of incubation has been shown to influence hemolytic reactions of beta-hemolytic streptococci. (6)

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
Streptococcus pyogenes
ATCC ® 19615
A 24hr 35°C CO 2 ** Growth; beta-hemolysis
Streptococcus pneumoniae
ATCC ® 6305
A 24hr 35°C CO 2 ** Growth; alpha-hemolysis
Staphylococcus aureus
ATCC ® 25923
A 24hr 35°C CO 2 ** Growth
Proteus mirabilis
ATCC ® 12453
B 24hr 35°C Aerobic Partial inhibition
Pseudomonas aeruginosa
ATCC ® 27853
B 24hr 35°C Aerobic Partial inhibition

User Quality Control

** Atmosphere of incubation is enriched with 5-10% CO 2 .

PHYSICAL APPEARANCE

Columbia CNA Agar should appear opaque, and cherry red in color.

S. pyogenes growing on Columbia CNA Agar

Streptococcus pyogenes (ATCC ® 19615) colonies growing on Columbia CNA Agar (Cat. no. A50). Incubated in CO 2 for 24 hours at 35ºC.

S. pneimoniae growing on Columbia CNA Agar

Streptococcus pneumoniae (ATCC ® 6305) colonies growing on Columbia CNA Agar (Cat. no. A50). Incubated in CO 2 for 24 hours at 35ºC.

S. aureus growing on Columbia CNA Agar

Staphylococcus aureus (ATCC ® 25923) colonies growing on Columbia CNA Agar (Cat. no. A50). Incubated aerobically for 24 hours at 35ºC.

Columbia CNA Agar

Uninoculated plate of Columbia CNA Agar (Cat. no. A50).

REFERENCES

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.

2. Tille, P., 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. Biochemical Tests for Identification of Medical Bacteria,, Lipincott Williams & Wilkins, Philadelphia, PA.

6. Jorgensen., 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.


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