Taxonomy
• Family: Micrococcaceae
• Genus:
– Staphylococcus
• Coagulase positive
• Coagulase negative
– Micrococcus
Laboratory Diagnosis:
Cultural Characteristics-
• Staphylococcus aureus
– Colony morphology
• Smooth, butyrous,
white to yellow,
creamy
• Grow well @ 18-24
hours
• S. aureus may produce
hemolysis on blood
agar
S. aureus
Genus Staphylococcus
• After gram staining if the results reveal
• gram-positive cocci
Staphylococcus spp streptococcus spp
Catalase Test
Staphylococcus spp streptococcus spp
Positive Negative
Identification Tests: Catalase
• Principle: tests for enzyme catalase
2 H2O2 2 H2O + O2
• Procedure
– Smear a colony of the organism to a slide
– Drop H2O2 onto smear
Catalase Test: Interpretation
– Presence of bubbles
• Positive
• Staphylococci
– Absence of bubbles
• Negative
• Streptococci
Genus Staphylococcus
Coagulase positive Coagulase negative
• S. aureus
• S. epidermidis
• S. saprophyticus
• S. haemolyticus
Coagulase is an enzyme like protein
Types of Coagulase
• 1. Bound Coagulase:
• Bound to the cell surfaceof bacteria.
• 2. Free Coagulase:
• Reside within the extracellular space.
Function of Coagulase
• 1. Causes plasma to clot by converting fibrinogen to fibrin
• 2. Its responsible for the pathogenicity of microorganism
Identification test: slide coagulase test
• Differentiates members within the Staphylococci
• Detects clumping factor found in S. aureus
• Detect the bound coagulase
• Procedure
– Place a drop of sterile water on a slide and emulsify a colony
– Add a drop of rabbit plasma to the suspension
– Observe
• Agglutination = Positive
• No agglutination= Negative
• Confirm negative by tube Coagulase Test
because bacteria may have free coagulase
Identification Tests: tube Coagulase Test
•Detects the extracellular enzyme
“free coagulase” or staphylocoagulase
•Causes a clot to form when bacterial cells are
incubated with plasma
•Procedure
• Inoculate plasma with organism and
incubate at 35-37 0 C
• Observe at 30 minutes for the presence of
a clot
• Continue for up to 4 hours, if needed
Novobiocin Susceptibility Test
• Test to differentiate coagulase-
negative staphylococci from
[Link] from urine
samples
– S. saprophyticus is resistant
(top)
– Other CNS are susceptible
Novobiocin sensitivity test
Distinguishing features of the major species of
staphylococcus
Characters [Link] [Link] [Link]
Coagulase + - -
Novobiocin Sensitive Sensitive Resistant
sensitivity
Acid from + - -
mannitol
fermentation
anaerobically
Phosphatase + + -
