A8 MYCOPLASMA AGAR

Technical Data #1726a / 2003.06.12

A8 Mycoplasma Agar is a solid medium used for detection and identification of Ureaplasma urelyticum and Mycoplasma sp. Fresh yeast extract, horse serum and enrichment permit luxuriant growth. Penicillin is the selective agent for the gram-positive bacteria.

FORMULA
in grams per liter purified filtered water

Casein Peptone	13,6
Soy Peptone	2,4
Potassium Phosphate, dibasic	2,0
Sodium Chloride	4,0
L-Cystein, HCI	0,1
Dextrose	2,0
GHL, tripeptide	0,00002
Calcium Chloride	0,14
Putrescine HCl	1,66
Urea	1,0
Agar, Purified	10,5
Supplements
Horse Serum	200 ml
Yeast Extract, fresh, pH 6,0	10 ml
Bio-X	5 ml
Antibiotic
Penicillin G	1,0 MM units

pH 6.0 +/- 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.

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).

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

1. Inoculate specimens such as urethral, vaginal, cervical swabs, urine sediment and others into Mycoplasma Transport Broth (#2281).
2. Transfer 0,1 ml of Mycoplasma-Transport Broth into each of the following media: U9 Broth (#2361), Arginine Broth (#2991) and Mycoplasma A7 (#1542) or A8 (#1726)
3. Incubate at 35º C with CO₂ or under anaerobic atmosphere for 3, 4, 7 days.
4. Examine agar plates at low power (100x).

QUALITY CONTROL
Results after 3 days at 25ºC anaerobic

Organisms	ATCC	Growth	Appearance
Ureaplasma urealyticum	27815	+	brown colonies
Ureaplasma urealyticum	27618	+	brown colonies
Mycoplasma hominis	23114	+	egg fried colonies

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

REFERENCES

1. Balows, A., W.J. Hausler 1981 Diagnostic Procedures for Bacterial Mycotic and Parasitic Infections, 6th ed.
2. 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, pp.407-411.
3. Mac Faddin, Jean F., Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria Vol.1 1985 Baltimore, MD. Williams & Wilkins.
4. Shepard, M.C., C.D. Lanceford 1976 Differential Agar Medium (A7) for identification of Ureaplasma urealyticum (human T-strain) in primary cultures of clinical material. J. Clin. Microbiol. 3:613-625.

CATALOGUE NUMBERS

Prepared media
Plates, 60x15 mm	10/pkg	1726
Tubes, 12x75 mm	10/pkg
Tubes, 13x100 mm	10/pkg	2989
Ureaplasma-Mycoplasma Transport Broth		2281
U9 Broth		2361
Arginine Broth		2991

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