Catalase Reagent: Formulation Per 100 ML

CATALASE REAGENT
- For in vitro use only - Catalogue No. RC35
Our Catalase Reagent is used to detect the To aid in species differentiation:

presence of the enzymes, catalase and peroxidase, 1. Aerococcus urinae (+) from Aerococcus
produced by some bacteria. viridans (−)
In the respiratory chain of all bacteria, 2. Pediococcus acidilactici (+) and P.
reduced flavoproteins and iron-sulfur proteins pentosaceus (+) from other Pediococcus (−)
unite with oxygen and oxidases to form two 3. Staphylococcus aureus subsp. anaerobius and
compounds, hydrogen peroxide and the S. saccharolytica (−) from other
superoxide radical. These compounds, if allowed Staphylococcus spp. (+)
to accumulate, are toxic to bacteria and results in 4. Campylobacter fetus, C. hyointestinalis, C.
their death. Bacterial survival is accomplished by jejuni, and C. coli (all +) from C. spurorum,
the production of specific enzymes that allow C. concisus, and C. mucosalis (all −)
bacteria to neutralize the toxic compound. 5. Neisseria elongata subsp. elongata, N.
Hydrogen peroxide is decomposed by the elongata subsp. nitroreducens, and N.
action of two enzymes: catalase and either a mucosa (all −) from other Neisseria spp. (+)
peroxidase, NADH, NADPH, cytochrome c, or 6. Moraxella bovis (variable) and Kingella spp.
glutathione. To observe the action of these (−) from other Moraxella spp. (+)
enzymes, catalase reagent, a dilute solution of 7. Prevotella oulorum (+) from other Prevotella
hydrogen peroxide, is added to a pure bacterial spp. (−)
culture. Any immediate bubbling is indicative of
a positive result since oxygen is a byproduct of
hydrogen peroxide decomposition. Formulation per 100 mL
Our catalase reagent can be used:
Hydrogen peroxide (3%) ........................ 100.0 mL
To differentiate between genera:
1. Streptococcus (−) from Micrococcus (+) and
Staphylococcus (V+) Recommended Procedure
2. Bacillus (+) from Clostridium (V−)
3. Listeria monocytogenes (+), Kurthia (+), A. Slide Method
Corynebacterium (+) from other
microorganisms that may be similar 1. With a sterile inoculating loop or wooden
morphologically: Erysipelothrix (−), applicator stick, pick the center a colony
Lactobacillus (V-), Enterococcus (−), and derived from an overnight culture plate and
group B Streptococcus (−) place it on a clean, glass slide.
4. Kingella denitrificans (−), Neisseria elongata 2. Place a drop of catalase reagent onto the
subsp. elongate (−), and Neisseria elongata smear.
subsp. nitroreducens (−) from other Neisseria 3. Observe for immediate bubbling. It may be
(+) and Moraxella catarrhalis (+) necessary to use a hand lens to detect weakly
5. Xenorhabdus spp. (−) from other positive reactions.
Enterobacteriaceae
6. Clostridium histolyticum, C. carnis, C.
tertium (all -) from other aerobic bacilli (+)
B. Tube Method Quality Control
1. Add 1.0-mL of Catalase Reagent directly to
Organism Expected Results
an overnight, heavily inoculated pure agar
slant culture. (Do not use a blood agar)
Staphylococcus aureus +ve Bubbling
2. Observe for immediate bubbling.
ATCC 25923
• For the slide method, do not reverse the Streptococcus -ve No bubbling
order of the procedure as platinum needles pyogenes
may produce false positive results. Nichrome ATCC 19615
wire does not cause bubbling
• Reactions are immediate although weak
reactions may be difficult to observe without Storage and Shelf Life
a hand lens
• The concentration of hydrogen peroxide in Our Catalase Reagent should be stored at 4
our Catalase Reagent is 3%; some catalase to 8°C and protected from light. Under these
test procedures require a 30% solution of conditions, the reagent has a shelf life of 26
hydrogen peroxide. Do not confuse the two weeks from the date of manufacture.
concentrations
• Colonies obtained from blood agar plates
should be picked and transferred very References
carefully. Any accidental transfer of the
medium to the slide will result in false 1. Mitchell RL, Anderson IC. Catalase
positive results since erythrocytes (RBCs) photoactivation. Science 1965; 150:74.
contain catalase/peroxidase. To avoid this 2. Funada H, Hattori K-I, Kosakai N.
problem chocolate agar can be used Catalase-negative Escherichia coli isolated
• Growth for catalase testing must be from an from blood. J Clin Micro 1978; 7:474-8.
18 to 24-hour culture since older colonies 3. Chester B, Moskowitz LB. Rapid catalase
may lose their catalase activity and yield supplemental test for identification of
false-negative results members of the family Enterobacteriaceae.
• Be aware that catalase-negative isolates may J Clin Micro 1987; 25:439-41.
exist for many species 4. Isenberg HD, Ed. Clinical microbiology
• When anaerobes are tested, cultures should procedures handbook, Vol I. Washington,
be exposed to air for 30 minutes prior to the DC: ASM, 1992.
addition of catalase reagent 5. Murray PR, Baron EJ, Pfaller MA, Tenover
• Catalase is inactivated by sunlight FC, Yolken RH. Manual of clinical
th
microbiology. 7 ed. Washington DC: ASM
Press, 1999.
Interpretation of Results 6. MacFaddin JF. Biochemical tests for
identification of medical bacteria. 3rd ed.
Positive: Immediate bubbling Philadelphia: Lippincott Williams &
(Oxygen is released) Wilkins, 2000.
Negative: No bubbling Original: April 2002

Revised / Revisited: January 2014

Catalase Reagent: Formulation Per 100 ML

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Catalase Reagent: Formulation Per 100 ML

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CATALASE REAGENT

- For in vitro use only - Catalogue No. RC35

Our Catalase Reagent is used to detect the To aid in species differentiation:

Negative: No bubbling Original: April 2002

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