Laboratory Evaluation of Platelets
Laboratory Evaluation of Platelets
ENUMERATION OF PLATELETS
- Counting of platelets is an important and logical starting point in evaluating bleeding problems
- Done by estimating the number of platelets on PBS or by manual or automated techniques
FONIO’S METHOD
- Indirect method that uses 14% MgSO4 as anticoagulant
SPECIMEN REQUIREMENTS
- Venous blood collected with EDTA
- Capillary puncture can yield low platelet counts secondary to platelet adhesion to the wound
o Capillary puncture should be performed with a larger blade so that a deep wound with a free
flowing blood is obtained
PLATELET FUNCTION
o ADHESION
Tests:
1. Bleeding Time Test
- Time it takes for a standard wound to stop bleeding
- Comprehensive test of platelet action in vivo
- Sensitive to abnormalities of platelet numbers and function to plasma VIII:vWF deficiencies
- Sensitive to abnormalities of vessel wall composition that interfere with platelet function
- Methods:
a. Duke Method
- Small cut in the earlobe is made and the length of time required for the bleeding to stop
is recorded
- NV: 1-3 minutes
b. Ivy Method
- Introduced standardization into the procedure and the time required to stop bleeding is
recorded
-
c. Mielke/Template BT
- Modified Ivy method
- Uses a template to produce a standardized slit in place of the disposable lancets
d. Simplate Method
- Uses a Simplate bleeding time device
- Uniform incision is made on the forearm and the length of time required for bleeding to
stop is recorded
o AGGREGATION
Performed to assist in the diagnosis of hereditary and acquired platelet disorders
Tests
1. Platelet Aggregation
- Measured with a platelet aggregometer
- Basic principle: Citrated, plasma-rich plasma is stirred in the aggregometer while a light beam
is passed through the suspension
- Chemical stimulus is added
ADP
Epinephrine
Collagen
Thrombin (retains aggregating properties but lacks clotting ability)
Ristocetin
Arachidonic acid
- The shape change from discoid to spheroid is monitored as an initial decrease in light
transmittance
- Subsequent aggregation allows an increase of light to pass through the suspension to the
photodetector and to be recorded as an increase in light transmittance
o PLATELET FACTOR ASSAYS
1. Platelet Factor 3 Availability
- Platelet factor 3 is a blood coagulation factor derived from platelets that act with certain plasma
thromboplastin factors to convert prothrombin to thrombin
- When platelets are incubated with kaolin and epinephrine, they are stimulated to provide PF3
activity
- The test compares the clotting time of the patient’s PRP with that obtained for a group of normal
individuals
- NV: 37 to 51 seconds for PRP
- Decreased in: Acquired or congenital defects of platelet secretion or release; impaired function
of factors V and Xa