PLATELET AGGREGOMETRY

Prepared by: Kenneth S. Destura

Principle:
Aggregating agents added to stirred suspension of platelet rich plasma (PRP)
induce a shape change and aggregation of platelets.
As the result, the PRP changes form a turbid suspension to one that transmits
more light as the aggregates are formed
hus, the process of platelet aggregation can be follo!ed by a platelet
aggregometer that measures a change in light transmission.
Aggregating Agents:
ADP
"pinephrine
#ollagen
hrombin
Ristocetin
Arachidonic acid
Special Patient Requirements:
Preferably, the patient and the normal control should be fasting. $ther!ise, they
should ha%e had a relati%ely fat&free meal' this eliminates the optical problem of
lipemic plasma.
he patient should not be ta(ing aspirin or nonsteroidal anti&in)ammatory drugs.
Specimen Requirements:
Platelet&rich plasma is obtained from both patient and control specimens by
centrifuging at R at *++ , g for -+ minutes.
As a conse.uence of processing of the PRP, platelets sometimes e,hibit a
/platelet shoc(0 phenomenon and !ill be poorly or not responsi%e to ADP,
collagen and epinephrine for as long as 1+ minutes. 2f a poor response is
obtained !ith one of these reagents, the test is repeated 1+ minutes later.
Procedure:
-. Perform platelet count on patient and control PRP' ad3ust the platelet counts to
*++ to 1++ , -+
4
56 by diluting !ith their respecti%e ali.uots of PPP. Keep samples
stoppered at room temperature.
*. 7arm +.8 m6 ali.uots of PRP in appropriate cu%ettes to 19# for se%eral minutes
as needed for testing.
1. Ad3ust -++: transmittance of the platelet aggregometer using patient PPP.
;. Place !armed PRP samples in the aggregometer and ad3ust the baseline reading
after at least - minute.
8. After stirring PRP for * minutes !ith small magnetic stir bars, forcefully add the
appropriate %olume of aggregating agent to ensure ade.uate mi,ing.
<. Record the aggregation cur%e for 1&8 minutes or until no further changed is
noted.
Repeat steps *&< for each aggregating agent !ith patient and control PRP.
Platelet aggregation patterns:
At optimal reagent concentrations, t!o aggregation !a%es are seen' the primary
response is follo!ed by a brief plateau, and then much larger secondary
response occurs during !hich platelets aggregate irre%ersibly.
=or the ma3ority of the samples, there is an optimal concentration of these
aggregating agents that results in t!o&!a%e (one&!a%e for collagen) response.
#ollagen produces a single !a%e of aggregation after a lag phase.
ADP
"pinephrine
hrombin
Ristocetin
- >?
-+ >?
+.1 units5m6
-.8 mg5m6
@#ollagen
Arachidonic acid
8 >g5m6
-.+ m?
Comments:
o ensure platelet %iability during the procedure, all testing should be completed
!ithin * hours of blood collection.
Specimens for platelet aggregation studies should not come in contact !ith any
type of glass!are. Alass !ill prematurely acti%ate platelets and thus result in a
less&than&optimal response to aggregating agents.
Specimens should not be refrigerated as platelets (ept cold do not respond
optimally to aggregating agents. Platelets are %iable longer if they are (ept at
room temperature and aggregation occurs best at 19#.
Because stirring eCects reagent mi,ing and platelet aggregate collisions, it has a
signiDcant eCect on platelet aggregation. ?ost aggregation instruments rotate
the stir bars at -*++ rpm.