CHOCOLATE AGAR

Technical Data #1299a / 2003.05.29

Chocolate Agar is an enriched medium for the isolation and cultivation of fastidious organisms as Neisseria and Haemophilus sp. Hemoglobin and Bio-X enrichment replace lysed heated blood and give to the culture media enough nutrients agents to permit luxuriant growth of fastidious microorganisms.

FORMULA
in grams per liter purified filtered water

Casein Peptone	7,5
Meat Peptone	7,5
Corn Starch	1,0
Potassium Phosphate, dibasic	4,0
Potassium Phosphate, monobasic	1,0
Sodium Chloride	5,0
Agar	10,0
Hemoglobin	10,0
Enrichment Bio-X	10 ml

pH 7,2 +/- 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 2-8º C protected from direct light and dehydrated powder, in a dry place, in tightly-sealed containers at 2-25º 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 36 g of dehydrated media in 500 ml of purified filtered water. Heat with frequent agitation and boil for one minute. Sterilize at 121º C for 15 minutes. Cool to 45-50º C. Add 500 ml of sterile 2% Hemoglobin (8662) and 10 ml of Bio-X Enrichment (8601). Mix gently and dispense into sterile Petri dishes or into steriles culture tubes.

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

1. Directly, inoculate plate as soon as possible with specimen and streak for isolation.
2. Incubate at 35º C with 5% CO₂ for 48 hours.

QUALITY CONTROL
Results after 48 hrs at 35ºC w/ CO₂

Organisms	ATCC	Growth
Neisseria gonorrheae	43069	+
Neisseria gonorrheae	clinical	+
Haemophilus influenzae	10211	+

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

REFERENCES

1. Lennette, E.H., Ballows, A., Hausler, W.J.Jr., and Shadomy, H.J. Manual of Clinical Microbiology. 4th ed. 1985 Washington D.C.: American society for Microbiology.
2. N.C.C.L.S. 1990 Quality Assurance for Commercially Prepared Microbiological Culture Media. Approved Standard. Vol.10, No.14. NCCLS Document M22-A.
3. Mac Faddin, Jean F., 1985 Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria. Vol.1. Baltimore, MD.: Williams & Wilkins.
4. Martin, J.E., T.E. Billings, J.F. Hackney, J.D. Thayer 1967 Primary isolation of N. gonorrheae with a new commercial medium. Public Health Rep. 82:361-363.

CATALOGUE NUMBERS

Dehydrated media	500 g	QB-39-1906
Hemoglobin	500 g	QB-39-2114
Prepared media
Plates, 100x15 mm	10/pkg	1299
Mono-Plates, 100x15 mm	10/pkg	1299M
Bi-plate: Choco/Thayer Martin, 100x15 mm	10/pkg	1330
Bi-plate: Choco/Martin Lewis, 100x15 mm	10/pkg	1457
Tubes, 20x125 mm	10/pkg	2929
Jembec Kit	10/pkg	1311
Bio-X Enrichment, 10 ml	10/pkg	8601
Hemoglobin, 2% sterile	500 ml	8662

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