COLUMBIA CNA AGAR 5% SHEEP BLOOD 6 mg VANCOMYCIN

Technical Data #1680a / 2000.11.01

Columbia CNA Agar 5% sheep blood with 6 mg of vancomycin is a selective solid medium for the isolation of Enterococcus faecalis Vancomycin Resistant. Antimicrobial agents, colistin and nalidixic acid, suppress growth of gram-negative organisms. Vancomycin inhibits Gram-positive microorganisms other than enterococci and enterococcus Vancomycin resistant.

FORMULA
in grams per liter purified filtered water

Casein Peptone	10,0
Meat Peptone	5,0
Yeast Extract	5,0
Beef Extract	3,0
Corn Starch	1,0
Sodium Chloride	5,0
Agar	13,5
Defibrinated Sheep Blood	50 ml
Antibiotics
Colistin	0,010
Nalidixic Acid	0,015
Vancomycin	0,006

pH 7,3 +/- 0,2 à 25º C

This approximate formula may be adjusted and/or enriched to obtain best results.

PRECAUTIONS
This medium is for IN VITRO diagnostic use only.

STORAGE
Store prepared media at 2-8° C protected from direct light and dehydrated powder, in a dry place, in tightly-sealed containers at 2-25° C.

SIGN OF DETERIORATION
Media should not be used if the expiry date has passed. Prepared media should not be used if there are signs of contamination or deterioration (shrinking, cracking, evaporation or discoloration). Do not use dehydrated media if it is caked.

DIRECTIONS
Suspend 42,5 g of dehydrated media in 950 ml of purified filtered water. Heat with frequent agitation and boil for one minute. Sterilize at 121° C for 15 minutes. Cool to 45-50° C. Aseptically add 50 ml of Defibrinated Sheep Blood (4245) and 2x5 ml of #8736 VRE Vancomycin Supplement. Mix thouroughly and dispense into Petri dishes.

PROCEDURE
Prior to inoculate, the prepared media should be brought to room temperature.

1. Directly, inoculate the specimen and streak for isolated colonies.
2. Incubate at 35° C for 24 hours.

QUALITY CONTROL
Results after 24 hrs at 35ºC

Organisms	ATCC	Growth
Enterococcus faecalis	29212	-
Enterococcus faecalis	51299	+
Enterococcus gallinarum	clinical	+

LIMITATIONS OF METHOD
This is only a part of the identification. Other tests may be required.

REFERENCES

1. Ballows, A., Hausler, W.J.Jr., K.L Herrmann, H.D. Isenberg, Shadomy, H.J. 1991 Manual of Clinical Microbiology. 5th ed. Washington D.C.: American Society for Microbiology.
2. Sahm, D.F., S. Boonlayangoor, P.C. Iwen, J.L. Baade, G.L. Woods 1991 Factors influencing determination of High-Level Aminoglycoside resistance in Enterococcus faecalis. J. Clin. Microbiol. 29:1934-1939
3. Swenson, J.M., N.C. Clark, M.J. Ferraro, D.F. Sahm, G. Doern, M.A. Pfaller, L.B. Reller, M.P. Weinstein, R.J. Zabransky, F.C. Tenover 1994 Development of a Standardized Screening Method for Detection of Vancomycin-Resistant Enterococci J. Clin. Microbiol. 32:1700-1704
4. Tenover, F.C., J. Tokars, J. Swenson, S. Paul, K. Spitalny, W. Jarvis 1993 Ability of clinical laboratories to detect antimicrobial agent-resistant enterococci J. Clin. Microbiol. 31:1695-1699.

CATALOGUE NUMBERS

Dehydrated media	500 grams	QB-39-1009
VRE Vancomycin Supplement	10 vials/pkg	8736
Defibrinated Sheep Blood	50 ml	4245
Prepared media
Plates, 100x15 mm	10/pkg	1680
Mono-Plates, 100x15 mm	10/pkg	1680M

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