COAGULASE TEST

Staphylococcus aureus

Because not all 'staph' are created equal!

 OUTLINE

Coagulase test Principle Laboratory

Slide Test Bound Coagulase Procedure
Tube Test Free Coagulase Results
Coagulase Test
Staphylococcus aureus

the ability to clot plasma continues to be the most widely used and generally
accepted criterion for the identification of pathogenic staphylococci
associated with acute infections

Slide (bound coagulase)

rapid screening, usually clot formation within four hours. If no clot formation is
observed during this period, perform a tube test.

Tube (free coagulase)

a definite examination that detects even the free coagulase. Need 24 hours
incubation of plasma with suspected colonies.
Principle
Coagulase Test

Coagulase is an enzyme-like protein and causes plasma to clot by converting

fibrinogen to fibrin. Staphylococcus aureus produces two forms of coagulase:
bound and free.

Bound coagulase (clumping factor)

is bound to the bacterial cell wall and reacts directly with fibrinogen. This results in an alternation
of fibrinogen so that it precipitates on the staphylococcal cell, causing the cells to clump when a
bacterial suspension is mixed with plasma. This doesn’t require a coagulase-reacting factor.

Free coagulase
involves the activation of plasma coagulase-reacting factor (CRP), which is a modified or
derived thrombin molecule, to form a coagulase-CRP complex. This complex in turn reacts
with fibrinogen to produce the fibrin clot.
Coagulase Test
Staphylococcus aureus
 Slide Coagulase Test
Staphylococcus aureus

With a wax pencil, divide the slide

into two. Label one side "C" for the
saline CONTROL and the other "T"
for the TEST.
Observe its hemolytic property on
BA and take note of its production
on TSA.
Place a small drop of NSS on the C side and
another drop on the T slide. Make heavy
suspension in these drops of NSS with
isolated colonies growing on BA.

Add a loopful of plasma in the TEST and mix

With the stock culture of thoroughly with a wire loop and observe for
Staphylococcus assigned to the the appearance of clumping within 10 seconds.
group, make a smear and do Gram Compare the test with the saline control.
staining.
 Tube Coagulase Test
Staphylococcus aureus

Get a small test tube containing 0.5

ml fresh human plasma.
Inoculate the plasma with a loopful of
bacteria from BA.
Observe its hemolytic property on
BA and take note of its production
on TSA.

Incubate at 37°C and examine for clotting at

intervals of 30 minutes for 4 hours. If no clot
is observed at the end of this period, examine
With the stock culture of the tube again at 6 and 24 hours. A small
Staphylococcus assigned to the amount of coagulum formed is weakly positive,
group, make a smear and do Gram a complete clotting is strongly positive.
staining.
 Results and Discussions
S. aureus

Slide Test Tube Test

(+) clumping (+) clumping

"STAPH! Look and Listen!"
 nks!
Tha