C.L.E.D. (CYSTINE LACTOSE ELECTROLYTES DEFICIENT) AGAR

Technical Data #1516a / 98.05.13

CLED Agar is a non selective solid medium for cultivation of pathogens from urine specimens. Lack of salts (electrolytes) inhibits swarming of Proteus sp. Bromthymol blue indicator differentiates bacteria between lactose fermenters and non lactose fermenters.

FORMULA
in grams per liter purified filtered water

Gelatin Peptone	4,0
Casein Peptone	4,0
Beef Extract	3,0
L-Cystine	0,128
Lactose	10,0
Bromthymol Blue	0,02
Agar	15,0

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 2-8º 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 1000 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. Mix gently and dispense into sterile Petri dishes.

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

1. Inoculate plates as soon as possible with urine specimen, using calibrated loop, streak for isolation.
2. Incubate at 35º C for 24 hours.

QUALITY CONTROL
Results after 24 hrs at 35º C

Organisms	ATCC	Growth	Colour
Escherichia coli	25922	+	yellow
Proteus mirabilis	12453	+	blue
Proteus vulgaris	8427	+	blue
Staphylococcus aureus	25923	+	yellow

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

REFERENCES

1. Dixon, J.M.S., M.A. Clark 1968 Experience with agar-filled spoons for the transport of specimens of urine. Can. Med, Assoc. J. 99:741
2. Mac Faddin, Jean F., 1985 Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria. Vol.1. Baltimore, MD.: Williams & Wilkins.
3. Mackey. J.P., G.H. Sandys 1966 Diagnosis of urinary tract infections Br. Med. J. 1:1173
4. Sandys, G.H. 1960 A new method of preventing swarming of Proteus spp with a description of a new medium suitable for use in routine laboratory practice. J. Med. Lab. Tecnol. 17:224

CATALOGUE NUMBERS

Dehydrated media	500 g	QB-39-1109
Prepared media
Plates, 100x15 mm	10/pkg	1516
Mono-Plates, 100x15 mm	10/pkg	1516M
Plates, 60x15 mm	10/pkg	1623
Plates, 150x15 mm	5/pkg	1495

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