  Some bacteria produce coagulase, an enzyme

and as a precursor of thrombin like substance

which coagulate plasma.
 There are two kinds of coagulase which could be
present in the microorganism – the free coagulase
and the bound coagulase.
 This enzyme is demonstrated in vitro by two
methods.
• The Test Tube Method
• The Slide Method
 Differentiate pathogenic from non-pathogenic
Staphylococcus
Tube Method:
The method measures free coagulase.

 Materials:
• Citrated or heparinized human or rabbit plasma is
diluted with isotonic saline or other suitable diluent
• Overnight broth culture of microorganism.

 Procedures:
Place about 0.5ml of diluted plasma in each of 2 test
tubes.
To one tube add 5 drops of an overnight broth culture of
the organism.
Do not place anything in the other tube which serves as
control
Incubate broth tube at 370C. Examine at 24 hours.
Result:

Positive Coagulase – a clot is formed

Negative Coagulase – no clot is
formed
Slide Method:
This usually detects bound coagulase, which is
the
clumping factor attached to the cells and acts directly
on
the fibrinogen.

 Procedures:
Divide the slide into two sections with grease pencil.
Place a small drop of normal saline on each area.
Emulsify one or two colonies of microorganism on
blood agar plate on each drop to make a smooth
suspension.
Add a drop of undiluted plasma to one of the drop of
saline and stir gently with a wire loop. Do not put
anything in the other drop that serves as control.
Observe for clumping of the microorganism