Clot Retraction Time

1. State the diagnostic importance of performing test for clot retraction.

⚫ Clot retraction - process by which a blood clot becomes smaller (shrinking of the blood), and
draws the edges of a broken blood vessel together (involves shortening of fibrin threads and
squeezing out of excess serum).
⚫ This test/process is important in order to know the promoting clot stability and maintaining
blood vessel patency
⚫ Helps in detecting the ability of the blood to clot formation.
⚫ Measured by % of serum yield
⚫ Influenced by: (a) number and activity of platelets, (b) concentration of fibrinogen, and (c)
acquired disorders of platelet function
⚫ % clot retraction: (Vol. of serum/Vol. of whole blood) x 100

2. Discuss the different methods used to determine clot retraction. Explain the procedure in clot
retraction
3.
Principle: A sample of blood in a calibrated tube is allowed to clot and the extend of retraction is
determined at 1,2,4 and 24 hours. This process depends on normal numbers of contractile platelets,
presence of calcium; ATP and a normal concentration of fibrinogen

Standard Procedure in Clot Retraction

1. Withdraw 3 mL of venous blood
2. Place 1 mL of blood into 3 glass test tubes, centrifuge tubes and immediately incubate in a
37°C water bath. Set a timer for 1 hour
3. At 1 hour, observe the clot and record results.
4. Inspect the tubes at 2, 4, and 24 hours, observe and record results
5. Examine the tubes for retraction after incubation
6. Separation of the clot from the test tube is complete retraction (4+) if clot retraction occurred
7. Measure the amount of serum expressed

METHODS: (refer to book of Ma’am Elsa page 254)

● Hirschboeck Method
Procedure:
1. one gtt of castor oil
2. one gtt of blood
3. start the timer and observe the dimpling to occur
Interpretation:
Normal Value: 15-45 minutes
<15 minutes - thrombotic tendency
> 15 minutes - hemorrhagic tendency
Incomplete or delayed occur in: - Thrombocytopenia and Glanzmann Thrombasthenia

NOTE: Blood will have a circular motion that resembles a dimple

- If dimpling occurs less than 15 minutes, you may have thrombotic tendency meaning it could form
unnecessary clots in your body
- If it is greater than 45 minutes, platelets are not enough or not functioning enough. You may have a
hemorrhagic tendency.
- This is a very simple test but not entirely reliable because it uses a small amount of blood

● Stefanini-Dameshek Method
Procedure:
1. Withdraw 3 mL of blood
2. transfer in a Wasserman tube
3. incubate at 37C or place at room temp allow 2 hours to elapse
4. after 2 hours have elapsed and observe the clot
Interpretation:
 Record observation:
no retraction
partial retraction
complete retraction
For appearance of the clot
Firm
Soft
NOTE:
- Makes use of a single test tube only unlike the standard method which uses 3 test tubes with 1
mL of blood
- Difference from original method is that this test uses only 1 test tube and is only observed for 2
hours
● Macfarlane Method
Procedure:
1. Place 5ml of fresh blood in a calibrated tube
2. Place a glass rod into the tube
3. Fit a cork at one end of the glass rod
4. Incubate at 37°C in a water bath
5. Examine every 5 -10 minutes for retraction
6. After coagulation, leave the tube at room temp for retraction
7. After retraction, remove the glass rod to remove clot
8. Measure the amount of serum left on the tube directly against the calibration tube
Interpretation:
Computation: % CR = mL of serum/ mL of blood x 100
Normal value: 44-67%
0 0 No serum extruded
1+ 5-10% serum extruded
2+ 10-20% serum extruded
3+ 20-35% serum extruded
4+ 35-50% serum extruded

● Tocantin’s Method
estimates extent of contraction by measuring size of clot in three dimensions from outside
of the tube, uses several small tubes
● Budtz- Olsen Method

4. Name the substance released by the platelet in order for the clot to retract.

• Actin, Myosin, & Thrombosthenin from the cytoplasm of platelets are resposinble for clot
retraction.
• Glycoprotein IIb/IIIa receptors are activated for platelets to bind with fibrin, & the
cytoplasmic proteins which causes clot retraction.
• Platelet integrin αIIbβ3
o mediates platelet response by rapidly transiting from its resting to an activated state
in which it serves as a receptor for ligands that can bridge platelets together

5. State the significance of an increased red cell fall-out.

Red cell fall-out - there is already a separation of the serum and the red blood cell
Increased red cell fall-out indicates decreased fibrinogen concentration
- Excessive RBC fall out indicates clot retraction but not firm
A. Fibrinogen concentration is low or absent
The red cell fallout is inversely proportional to the amount of fibrinogen present in
blood
- Fibrinogen will be converted to fibrin to hold the clot but it is low or absent
- Fibrinogen helps RBCs to be intact and firm so that there will be clot retraction
B. A fibrinolysin may be present
 ➔ The red cell fallout is directly proportional to the amount of fibrinolysin present
➔ Degrades fibrin or clot that has formed
C. Factor XIII deficiency will result in excessive red cell fallout
➔ Factor XIII is activated by thrombin and by strengthening the bonds between the
fibrin threads, produces an insoluble clot
➔ Factor XIII is a fibrin stabilizing factor
D. Budzt Olsen Method
➔ Increased rbc fall out at the bottom of the tube
- Indicates a slightly decreased of fibrinogen and should be noted on the result.
➔ Clot retraction is inversely proportional with the plasma fibrinogen concentration
- High RBC count limits the degree of retraction because of large volume of redcell within
clot.

6. Identify the standard reference values for clot retraction.

Normal Clot retraction - occurred 2-4 hours

Poor Clot retraction - occurs after 4 hours, and 24 hours
None NO RETRACTION - occurs after 24 hours

7. Identify potential sources of error when performing clot retraction.

⚫ Shaking or jarring of the test tube (falsely shortened CRT)
⚫ Temperature (increasing temp accelerates clot formation but weakens clot)
⚫ Faulty venipuncture or probing while drawing out blood.
⚫ Certain anemic patients with a low hematocrit value show increased clot retraction due to the
formation of a small clot.

8. Identify the conditions that may yield abnormal clot retraction.

⚫ Fibrinogen deficiency (congenital or acquired) = prolonged CRT
⚫ Thrombocytopenia (less than 100,000) = prolonged CRT
⚫ Thrombosthenia. Platelet would not function properly. Platelets could not pull or firm the fibrin
therefore leading to prolonged or no CRT
⚫ Polycythaemia vera = prolonged CRT Red blood cells are needed to form a clot.
If too much RBCs in the clot,platelets will have a hard time to shrinking the clot
leading to prolonged CRT Unlike in anemia, where clot retraction is increased / shorten
because you have red cells present in the body to remove the clot since during the set up dira
ga bind ang clot. (F XII) Prolonged, because RBC count limits the degree of clot retraction
⚫ Reduced clot formation:
⚫ Glanzmann thrombasthenia, DIC, hypofibrinogenemia, dysfibrinogenemia (small clot with
increased redblood cell “fall-out”) = prolonged CRT
⚫ DIC = prolonged
⚫ Hypofibrinogenemia = prolonged
⚫ Dysfibrinogenemia = prolonged
⚫ Disturbances in the coagulation system: (leads to thrombin formation and thus reduced platelet
activation) (Factor 7,8,11,13, and fibrinogen) - decreased clot retraction.
⚫ Glanzmann thrombasthenia: reduced glycoprotein IIb / IIIa causes reduced platelet aggregation
and clot retraction
Clinical SIgnificance: (PPT)

• Clot retraction is directly proportional to platelets, & inversely proportional with plasma
fibrinogen concentration.
• Measures coagulation ability.
• Bernard-Soulier Syndrome
• Measuring Hemostatic therapy.
• Excessive bleeding.

Reference:

Juanillo, E. F., Mario, J. P., Visitacion, R. P., & Perez, J. G. (2016). Essentials of Clinical
Hematology: A Study Guide for Students. Moake, J. L. (2023, April 18).

Bernadette F. Rodak. George A. Fritsma, Kathryn Doig. (2007). Hematology : clinical

principles and applications. St. Louis, MO :Saunders Elsevier