PRODUCT DETERIORATION This product should not be used if (1) after rehydration, the color of the broth is not light-orange but rather pink to red, (2) the expiration date has passed, or (3) there are other signs of deterioration. RAPID BROTH (LYOPHILIZED) SPECIMEN COLLECTION, STORAGE, TRANSPORT Specimens should be collected and handled following INTENDED USE recommended guidelines.4,5 Remel Rapid Urea Broth (Lyophilized) is recommended for use in qualitative procedures for rapid differentiation MATERIALS REQUIRED BUT NOT SUPPLIED of microorganisms on the basis of production. (1) Loop sterilization device, (2) Inoculating loop, swabs,

collection containers, (3) Incubators, alternative SUMMARY AND EXPLANATION environmental systems, (4) Supplemental media, The detection of urease activity is routinely used in (5) Quality control organisms, (6) Saline or demineralized clinical microbiology laboratories to aid in identification water, (7) Plastic tubes, (8) Water bath, (9) Pipettes. of gram-negative bacteria. In 1941, Rustigian and Stuart developed a medium for detection of urease PROCEDURE activity by Proteus spp.1 In further testing, they 1. Reconstitute Rapid Urea Broth (Lyophilized) by determined other urea-positive genera within the adding 3 ml of demineralized water or neutral Enterobacteriaceae could be detected in a medium with saline (pH 7.0 ± 0.2) to the vial. Mix to dissolve. a reduced buffering capacity.2 Rapid Urea Broth is such The color of the broth should be light-orange. If the a medium. It allows for rapid urease detection in enteric color is pink or red, do not use. gram-negative bacilli other than Proteus spp. This 2. Inoculate the 3 ml vial directly or aliquot 0.5 ml of medium is recommended by the American Public Health Association.3 the broth into plastic tubes. The inoculum may be removed from TSI Agar, KIA, Tryptic Soy Agar, or other agar medium having heavy growth in pure PRINCIPLE culture. To obtain rapid results, use a large Microorganisms that hydrolyze urea by means of the inoculum, 2-3 loopfuls of the test isolate. urease enzyme produce ammonia. This alkaline end 3. Incubate the tube aerobically at 33-37°C. The use product causes the pH indicator, phenol red, to change of a water bath will accelerate results. from yellow-orange to pink-red. A balance of 4. Examine for a pink-red color development at 15 monopotassium phosphate and disodium phosphate minutes, 30 minutes, 60 minutes and 4 hours. comprise the buffering system to produce a rapid change in pH, changing phenol red indicator accordingly. Note: Store rehydrated broth at 2-8°C and use on the day it is prepared.6 REAGENTS (CLASSICAL FORMULA)* Urea ...... 20.0 g INTERPRETATION Yeast Extract...... 0.1 g Positive Test - A pink-red color development Phenol Red ...... 0.01 g Negative Test - No color change Disodium Phosphate...... 0.095 g

Monopotassium Phosphate...... 0.091 g Demineralized Water ...... 1000.0 ml QUALITY CONTROL All lot numbers of Rapid Urea Broth (Lyophilized) have pH 6.9 ± 0.2 @ 25°C been tested using the following quality control organisms

*Adjusted as required to meet performance standards. and found to be acceptable. Positive and negative controls should be tested in accordance with established PRECAUTIONS laboratory quality control procedures. If aberrant quality This product is for In Vitro diagnostic use and should be control results are noted, patient results should not be used by properly trained individuals. Precautions should reported. be taken against the dangers of microbiological hazards by properly sterilizing specimens, containers, and media CONTROL INCUBATION RESULTS after use. Directions should be read and followed Cryptococcus neoformans Aerobic, up to Positive carefully. ATCC® 34877 4 h @ 33-37C

Proteus mirabilis Aerobic, up to Positive STORAGE ® ATCC 12453 4 h @ 33-37C Store lyophilized product in its original container at 2-8°C until used. Allow product to equilibrate to room Aerobic, up to Negative temperature before use. ATCC® 25922 4 h @ 33-37C LIMITATIONS PACKAGING 1. The use of glass tubes may result in false-positive Rapid Urea Broth (Lyophilized): results and is not recommended. REF R20388, 3 ml/Vial ...... Each 2. Urea test media rely on demonstration of alkalinity and are not specific for detection of urease activity. Symbol Legend Peptones in the medium may be hydrolyzed releasing amino acids which raise the pH and may REF Catalog Number result in false-positive reactions. A control test using the same test medium without urea can be IVD In Vitro Diagnostic Medical Device used to facilitate interpretation of questionable reactions.6 LAB For Laboratory Use

BIBLIOGRAPHY Consult Instructions for Use (IFU) 1. Rustigian, R. and C. Stuart. 1941. Proc. Soc. Exp. Biol. Med. 47:108-112. Temperature Limitation (Storage Temp.) 2. Rustigian, R. and C. Stuart. 1945. J. Bacteriol. 49:419- 445. LOT Batch Code (Lot Number) 3. Food and Drug Administration. 2000. Bacteriological Analytical Manual Online. Chapter 5, Updated Dec. Use By (Expiration Date) 2007. Authors: Andrews, W.H. and T.S. Hammack. AOAC International, Gaithersburg, MD. 4. Murray, P.R., E.J. Baron, J.H. Jorgensen, M.L. Landry, ATCC is a registered trademark of American Type Culture Collection. and M.A. Pfaller. 2007. Manual of Clinical Microbiology. 9th ed. ASM Press, Washington, D.C.

5. Forbes, B.A., D.F. Sahm, and A.S. Weissfeld. 2007. th Bailey and Scott’s . 12 ed. Mosby Elsevier, St. Louis, MO. 6. MacFaddin, J.F. 2000. Biochemical Tests for Identification of Medical Bacteria. 3rd ed. Lippincott IFU 20388, Revised September 1, 2009 Printed in the U.S.A. Williams & Wilkins, Philadelphia, PA.

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