Biochemical methods for

Staphylococci Identification
Catalase test
• What is catalase?
> Catalase is an enzyme produce by mostly aerobic
bacteria and facultative anaerobic bacteria.
> Catalase converts H2O2> into Water and Oxygen
neutralizing the effect of H2O2 to the bacteria
increasing the virulence factor and viability of the
bacteria
Catalase test

• Facilitates the detection of catalase enzyme in the

bacteria.
• It is essential for differentiating catalase-positive
micrococcaceae, staphylococcus from catalase
negative streptococcus in short staph and micrococcus
vs strep.
Catalase test
The principle behind this test is that catalase converts 3%
hydrogen peroxide into water and oxygen

For interpretation:
Rapid continuous bubbles(effervescence) : positive(+)
Lack of bubble formation after 30 seconds: NEGATIVE(-)
(+): Staphylococcus aureus
(-): Streptococcis pyogenes
Coagulase test
The coagulase test differentiates the strains of
Staphylococcus aureus from other Staph negative or
what we call the (CONS) Coagulase negative
staphylococcus
Example of CONS
Staphylococcus epidermidis
Staphylococcus saphrophyticus
Coagulase test
The principle behind this test is that the coagulase converts soluble
fibrinogen into soluble fibrin.
Slide test is screening test.
-it detects bound coagulase( clumping factor)
- of negative tube test will be performed.
Tube test.
- detects free coagulase
Coagulase test
• Interpretation:
• Positive results: white fibrin clots in plasma
ex. Staphylococcus aureus
• Negative results: Smooth suspension
ex. Staphylococcus epidermidis
Slide agglutination test
• It is solely for the identification of Staphylococcus
aureus.
• Agglutination of latex bead coated with fibrinogen
and antibodies to protein A ( protein in cell wall of S.
aureus)
Novobiocin test
• Novobiocin test is for the differentiation of
Staphylococcus saphrophyticus from other Coagulase
negative staphylococcus( CONS)
• Principle behind this test is we used antibiotic to
determine the susceptibility and resistability of the
bacteria.
• Need a 5ug novobiocin
Novobiocin test
Interpretation:
Susceptible: have a zone diameter of > 16mm
ex.staph epidemidis
Resistant: less than or equal to 16mm
ex. Staph saphrophyticus
Dnase test
• Detects Dnase activity in aerobic bacteria
• Dnase medium using methyl green becomes colorless in the
prescence of dnase.
• Positive: hydrolysis of the surrounding medium resulting in a
clear zone
ex. Staph aureus
• NEGATIVE: no clearing is observed
ex. Staph epidermidis
Mannitol Salt Agar
Selective/differential medium for staphylococcus species
Staphylococcus species are able to grow in 7.5% salt.
S. Aureus ferments mannitol turning phenol-red into yellow
Growth without fermentation plate remains pink to red with
no yellow halo surrounding growth
Growth with fermentation yellow halos surrounding growth.