Blood Coagulation
Part 2
Steps of blood coagulation
• There are three steps of the coagulation
process:
• Vascular spasm(Vessel constriction)
• The formation of a platelet plug
• Coagulation (blood clotting)
Prothrombin Time (PT)
• -Prothrombin time (PT) is a blood test that measures
how long it takes blood plasma to clot.
• - It is a measurement that assesses the extrinsic and
common pathways of the coagulation cascade (factors
VII, V, X, prothrombin, and fibrinogen).
• - Each laboratory has its own normal value, usually
between 11 and 15 seconds.
• PT test Procedure:
• 1. Collect blood samlpe in sodium citrate tube.
• 2. To collect plasma, centrifuge anticoagulated blood at 2,500 rpm for 10
minutes as soon as possible after collection.
• 3- In a tube, place 100 ul of plasma. Incubate for 1 minute in a water bath
at 37ºC.
• 4- start stopwatch
• 5- Add 200 ul pre-heated PT reagent (rabbit brain thromboplastin with 10
mM calcium chloride final concentration).
• 6- Prior to the estimated coagulation time, remove the tube from bath,
gently sliding the liquid content from the bottom to the middle of the tube
and stop the stopwatch at the time of clot appearance.
• 7- Record clot formation time.
Partial Thromboplastin Time (PTT)
• Partial thromboplastin time (PTT) is a blood
test that looks at how long it takes for blood
plasma to clot.
• The PTT test is used to measure and evaluate
all the clotting factors of the intrinsic and
common pathways of the clotting cascade.
• The PTT evaluates factors I (fibrinogen), II
(prothrombin), V, VIII, IX, X, XI and XII.
• Normal range is between 30 to 40 seconds.
• PTT test procedure
• 1. Collect blood samlpe in sodium citrate tube.
• 2. Centrifuge anticoagulated blood at 2,500 rpm for 10 minutes as soon as possible after collection.
• 3. Incubate a sufficient amount of 0.025M CaCl2 at 37 C.
• 4. Pipette 100 ul of plasma into test tube.
• 5. Pipette 100 ul ml of PTT reagent ( kaolin (hydrated aluminum silicate) and cephalin. Kaolin serves
to activate the contact-dependent Factor XII, and cephalin substitutes for platelet phospholipids) into
the test tube containing the plasma.
• 6. Mix the contents of the tube quickly and place in a 37 C water bath for 5 minutes.
• 7. After exactly 5 minutes, forcibly inject 100 ul of the prewarmed CaCl2 and simultaneously start the
stopwatch. Mix the test tube immediately and allow the test tube to remain in the water bath, gently
tilting the tube every 5 seconds.
• 8. At the end of 20 seconds, remove the test tube from the water bath . Hold the tube so that
contents can be monitored for the formation of the clot. Gently tilt the tube back and forth until a clot
forms, at which point, the timing is immediately stopped.
PTT test procedure
Citrated plasma, an activating agent, and
phospholipid are added together and
incubated at 37°C. Calcium is added, and the
time necessary for the clumping of kaolin is
measured.
EXAMPLE OF CONDITIONS THAT
PT RESULT PTT RESULT
MAY BE PRESENT
Liver disease, vitamin K insufficiency,
decreased or defective factor VII,
chronic low-grade
Prolonged Normal
disseminated intravascular coagulatio
n (DIC)
, anticoagulation drug (warfarin)
therapy
Decreased or defective factor VIII, IX,
XI, or XII, von Willebrand disease
(severe type), presence of
Normal Prolonged
lupus anticoagulant, autoantibody
against a specific factor (e.g., factor
VIII)
Decreased or defective factor I, II, V
Prolonged Prolonged or X, severe liver disease, acute DIC,
warfarin overdose
May indicate normal hemostasis;
however, PT and PTT can be normal
in conditions such as mild
deficiencies in coagulation factor(s),
Normal Normal or slightly prolonged mild form of von Willebrand disease,
and presence of weak lupus
anticoagulant. Further testing may be
required to diagnose these
conditions.
Good Luck
