UREA AGAR BASE
INTENDED USE
Remel Urea Agar Base is a solid medium recommended for use in qualitative procedures for the differentiation of microorganisms on the basis
of urease activity.
SUMMARY AND EXPLANATION
This medium was developed by Christensen to aid in the differentiation of rapid urea-positive Proteus spp. from other urea-positive enteric
1,2
gram-negative bacilli. Gelatin peptone and dextrose promote rapid growth of many enteric gram-negative bacilli. This allows for urease
3
activity to be demonstrated in organisms with more complex nitrogen requirements, such as Klebsiella and Enterobacter. This medium may
also be used to determine urease activity of nonenteric gram-negative bacilli, such as Brucella and Bordetella, and to differentiate
4,5
Cryptococcus from other yeasts.
PRINCIPLE
Urea Agar is a lightly buffered medium containing urea and phenol red, a pH indicator. When organisms utilize urea, ammonia is formed
which turns the medium alkaline. The indicator, phenol red, changes the medium color from pale-yellow to pink-red in an alkaline
environment. Gelatin peptone promotes rapid growth of enteric gram-negative bacilli, permitting a decrease in incubation time. Dextrose
stimulates urease activity in those organisms which hydrolyze urea slowly.
REAGENTS (CLASSICAL FORMULA)*
Urea ............................................................................ 20.0 g Dextrose ....................................................................... 1.0 g
Sodium Chloride............................................................ 5.0 g Gelatin Peptone............................................................ 1.0 g
Monopotassium Phosphate........................................... 2.0 g Phenol Red................................................................. 12.0 mg
pH 6.8 ± 0.2 @ 25°C
*Adjusted as required to meet performance standards.
PRECAUTIONS
This product is For Laboratory Use only. It is not intended for use in the diagnosis of disease or other conditions.
PREPARATION OF DEHYDRATED CULTURE MEDIUM
1. Suspend 29 g of Urea Agar Base in 100 ml of demineralized water.
2. Mix thoroughly and sterilize by filtration.
3. Dissolve 15 g of agar (REF R451011) in 900 ml of demineralized water.
4. Sterilize by autoclaving at 121°C for 15 minutes.
5. Cool to 45-50°C and aseptically add the prepared sterile Urea Agar Base.
6. Mix thoroughly and dispense into sterile tubes.
7. Cool in a slanted position so that deep butts are formed.
PROCEDURE
1. Consult current editions of appropriate references for the recommended procedure for sample preparation, inoculation, testing, and
interpretation.
INTERPRETATION OF THE TEST
Positive Test - Intense pink-red color development
Negative Test - No color change
QUALITY CONTROL
Each lot number of Urea Agar Base has been manufactured, packaged, and processed in accordance with current Good Manufacturing
Practice regulations. All lot numbers have been tested using the following quality control organisms and have been found to be acceptable.
Testing of control organisms should be performed in accordance with established laboratory quality control procedures. If aberrant quality
control results are noted, sample results should not be reported.
CONTROL INCUBATION RESULTS
®
Escherichia coli ATCC 25922 Ambient, 18-24 h @ 33-37°C Negative
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Klebsiella pneumoniae ATCC 27736 Ambient, 18-24 h @ 33-37°C Positive
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Proteus vulgaris ATCC 8427 Ambient, 18-24 h @ 33-37°C Positive
LIMITATIONS
1. Urea test media rely on demonstration of alkalinity and are not specific for detection of urease activity. Peptones in the media may be
hydrolyzed releasing amino acid residues, raising the pH, and resulting in false-positive reactions. A control test using the same test
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medium without urea can be used to facilitate interpretation of questionable reactions.
2. For Urea Agar results to be valid for rapid urease-positive organisms (e.g., Proteus spp.), the slant should be read within the first 2-6
3
hours after inoculation/incubation. Other members of the Enterobacteriaceae, such as Klebsiella and Enterobacter, exhibit a delayed
urease reaction which may require 24-48 hours of incubation.
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BIBLIOGRAPHY
1. Christensen, W.B. 1946. J. Bacteriol. 52:461-466.
th
2. Ewing, W.H. 1986. Identification of Enterobacteriaceae. 4 ed. Elsevier, New York, NY.
rd
3. MacFaddin, J.F. 2000. Biochemical Tests for Identification of Medical Bacteria. 3 ed. Lippincott Williams & Wilkins, Philadelphia, PA.
4. MacFaddin, J.F. 1985. Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria. Vol. 1. Williams & Wilkins,
Baltimore, MD.
th
5. Larone, D.H. 2002. Medically Important Fungi, A Guide to Identification. 4 ed. ASM Press, Washington, D.C.
Refer to the front of Remel Technical Manual of Microbiological Media for General Information regarding precautions, product storage and
deterioration, sample collection, storage and transportation, materials required, quality control, and limitations.
ATCC® is a registered trademark of American Type Culture Collection
IFU 455201, Revised February 15, 2011 Printed in U.S.A.
12076 Santa Fe Drive, Lenexa, KS 66215, USA
General Information: (800) 255-6730 Website: www.remel.com Email:
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