CATALASE REAGENT

Cat. no. Z62 Catalase Reagent, 3% 15ml
Cat. no. Z76 Catalase Reagent, 3% 60ml
Cat. no. Z262 Catalase Reagent, 15% 15ml

INTENDED USE

Hardy Diagnostics' Catalase Reagents are useful in the presumptive identification and differentiation of many bacteria. Beta-hemolytic organisms, such as Streptococcus species (catalase-negative), Staphylococcus species (catalase-positive), and Listeria species (catalase-positive) can be differentiated by their catalase reaction using 3% hydrogen peroxide. For catalase testing of anaerobic bacteria, 15% hydrogen peroxide appears to be more sensitive than 3% hydrogen peroxide.

SUMMARY

Most cytochrome containing organisms produce a catalase enzyme which breaks down hydrogen peroxide into oxygen and water. When a small amount of a catalase producing organism is introduced into hydrogen peroxide, bubbles of oxygen form as a result of the enzyme's activity.

REAGENT FORMULA

Z91 and Z76 include:

Hydrogen Peroxide, 3%, and Acetophenetidine.

Z262 includes:

Hydrogen Peroxide, 15% and Acetophenetidine.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-30ºC away from direct light. Products should not be used if there are any signs of deterioration (shrinking, cracking, or discoloration), contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat and freezing.

PRECAUTIONS

PROCEDURE

Specimen Collection: These products are not intended for primary isolation of patient specimens. These products are used in conjunction with other biochemical tests to identify cultures of isolated organism.

Method of Use:

Catalase Reagent, 3%:

With a loop or sterile wooden stick, transfer a small amount of a well isolated, 18 to 24 hour old colony from a non-blood-containing agar onto the surface of a clean, dry, glass slide. Immediately place a drop of Catalase Reagent, 3% onto a portion of the colony on the slide. Do not introduce a metallic loop into the drop, because this often causes a false-positive reaction.

Catalase Reagent, 15%:

Touch the center of a well isolated, 24 to 72 hour old colony with a non-metallic-loop or sterile wooden stick; avoid contact with the agar. Transfer growth onto the surface of a clean, dry, glass slide. Immediately place one drop of Catalase Reagent, 15% onto the smear. Do not introduce a metallic loop into the drop, because this often causes a false-positive reaction.

INTERPRETATION OF RESULTS

Observe for the immediate evolution of gas bubbles indicating a positive test. Formation of rare bubbles after 20 to 30 seconds is considered a negative catalase test.

LIMITATIONS

Aerobic organisms must be taken from an 18 to 24 hour old culture. Organisms lose their catalase activity with age. For slower growing anaerobic organisms, an older (24-72 hours) culture may be acceptable. Anaerobic cultures should be exposed to ambient air for a minumum of 30 minutes before testing.

It is recommended that colonies to be tested with the Catalase Test be taken from non-blood containing media due to the endogenous catalase activity present in animal red blood cells. For genera that require blood-containing media for growth, use isolates from a nonselective blood agar and avoid touching the agar with the loop.

Do not introduce a metallic loop into the drop of Catalase Reagent, because this often causes a false-positive reaction.

Bacteria grown on media with low levels or no glucose may yield conflicting results from pseudocatalase, a non-iron enzyme. The pseudocatalase reaction can be prevented by using media with 1% glucose.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, microscope slides, incinerators, incubators, pasteur pipets, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Reaction
Catalase Reagent, 3% (Cat. no. Z62 and Z76):
Staphylococcus aureus
ATCC ® 25923
Positive, bubbles seen
Streptococcus pyogenes
ATCC ® 19615
Negative, no bubbles seen
Catalase Reagent, 15% (Cat. no. Z262):
Bacteroides fragilis
ATCC ® 25285
Positive, bubbles seen
Clostridium perfringens
ATCC ® 13124
Negative, no bubbles seen

User Quality Control

PHYSICAL APPEARANCE

Catalase Reagent should appear clear and colorless.

Positive Catalase Test

Showing positive catalase test
Growth from a 24 hour Staphylococcus aureus (ATCC ® 25923) culture on TSA (Cat. no. G60) was applied to a sterile slide. A drop of Catalase Reagent (Cat. no. Z62) was dropped onto the applied growth. The production of gas bubbles was indicative of a positive catalase reaction. Catalase tests should not be performed from growth cultured on a blood-containing medium.

Negative Catalase Test

Showing negative catalase test
Growth from a 24 hour Streptococcus pyogenes (ATCC ® 19615) culture on TSA (Cat. no. G60) was applied to a sterile slide. A drop of Catalase Reagent (Cat. no. Z62) was dropped onto the applied growth. The absense of gas bubbles was indicative of a negative catalase reaction. Catalase tests should not be performed from growth cultured on a blood-containing medium.

REFERENCES

1. Versalovic, J., et al. Manual of Clinical Microbiology. American Society for Microbiology, Washington, D.C.

2. Tille, P.M., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

3. Anderson, N.L., et al. Cumitech 3B; Quality Systems in the Clinical Microbiology Laboratory, Coordinating ed., A.S. Weissfeld. American Society for Microbiology, Washington, D.C.

4. Isenberg, H.D. Clinical Microbiology Procedures Handbook , Vol. I, II & III. American Society for Microbiology, Washington, D.C.

5. Koneman, E.W., et al. Color Atlas and Textbook of Diagnostic Microbiology. J.B. Lippincott Company, Philadelphia, PA.

6. MacFaddin, J.F. 1985. Media for Isolation, Cultivation, Identification, Maintenance of Bacteria , Vol. I. Williams & Wilkins, Baltimore, MD.

7. Centers for Medicare and Medicaid, Appendix C, Survey Procedures and Interpretive Guidelines for Laboratories and Laboratory Services . Subpart K - Quality System for Non-Waived Testing. 493;1200-1265. www.cms.hhs.gov/clia .


ATCC is a registered trademark of the American Type Culture Collection.

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