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Platelet Function Testing: PFA-100

Introduction
The PFA-100 is a system for analysing platelet function in which citrated whole blood is
aspirated at high shear rates through disposable cartridges containing an aperture
within a membrane coated with either collagen and epinephrine (CEP! or collagen and
A"P (CA"P!# These agonists induce platelet adhesion$ acti%ation and aggregation
leading to rapid occlusion of the aperture and cessation of blood &ow termed the closure
time (CT!#
Advantages of the PFA-100 include:
'nly small %olumes of citrated %enous blood ()00*+! are needed and so the
test is useful for in%estigating platelet function in children#
Can be used by non-s,illed personnel and is both rapid and automated
The PFA-100 was designed as a screen to detect problems with primary
haemostasis and in part to replace the bleeding time and in this respect it is
better standardised#
-easurement of platelet function at high shear (physiological. rates whereas
+TA measures platelet function at low shear rates i#e# less physiological#
/elati%ely insensiti%e to clotting factor de0ciencies
1igh negati%e predicti%e %alue 2 i#e# if the PFA-100 gi%es a normal result then
with some e3ceptions primary haemostasis is intact (E3ceptions4 5P"$
Primary 5ecretion "efects$ mild Type 1 67".

A number of %ariables ha%e been shown to a8ect the results obtained with the PFA-100
and these include4
Variable !ect
Citrate
concentration
+aboratories must use a 03ed citrate concentration
Collection time 1ow the sample was collected and transported to the lab#
The tests must be performed within 9 hours of collection
1aematocrit Closure times increase progressi%ely with decreases in
haematocrit# Con%ersely$ CTs are shortened in the neonate
due to their higher haematocrit#
Platelet count Closure times increase progressi%ely as the platelet counts
falls below 100 3 10
:
;+
<lood =roup and
67F le%els
Closure times correlate in%ersely with plasma 67F acti%ity
le%els and may be increased in blood group ' patients for
the same reason (<lood group ' indi%iduals ha%e lower
67F le%els#.
"rugs4 C'> inhibitors such as aspirin and ?5A"s usually prolong
the closure time of the CEP cartridge but not the CA"P
cartridge#
The e8ects of A"P receptor bloc,ers such as Clopidogrel is
unpredictable#
nhibition of the =pb;a receptor is associated with a
signi0cant prolongation of both cartridges#
Ac@uired platelet
function defects
Cardio-pulmonary bypass
+i%er disease
Araemia
5ome foods =eneration of false positi%es particularly with the CEP
cartridge# This is due probably to ingested foods or drugs#
An abnormal PFA-100 result is not$ therefore$ diagnostic#
As"irin #esistance: The PFA-100 is often used to establish the presence of absence of
aspirin resistance# The fre@uency of aspirin resistance is un,nown$ but estimates range
from B-C0D# The mechanism of aspirin resistance is un,nown but proposed mechanisms
include poor patient compliance$ poor aspirin absorption$ increased platelet
hypersensiti%ity to agonists$ increased C'> acti%ity$ and polymorphisms in the =p a
receptor and the C'> enEyme# Aspirin resistance appears to be dose related in some
patients and may be o%ercome with higher doses#
Inter"retation
The following table summarises some of the abnormalities that ha%e been reported with
the PFA-100#
$isorder %T %ollagen-A$P %T %ollagen-PI
?ormal ? ?
Aspirin and ?5A"s ? F
A"P receptor disorders including the
use of Clopidogrel
? or F ? or F
<55 F F
=TT F F
67" F F
Platelet-Type 67" F F
"ense =ranule "e0ciency ? or F ? or F
Primary 5ecretion "efects ? or F ? or F
=ray Platelet 5yndrome F F
-G1:-related "isorders ? F
5cott 5yndrome ? ?
-"5 ? or F ? or F
+i%er "isease F (possibly as a
result of H1b.
F (possibly as a
result of H1b.
Araemia F (possibly as a
result of H1b.
F (possibly as a
result of H1b.
#eference #anges
/eported reference ranges for closure times are4
1 I) - 1:: seconds for the CEP cartridge
J BB - 1KI seconds for the CA"P cartridge
&
'hat Test Ne(t
1# The PFA-100 has a high negati%e predicti%e %alue i#e# if the PFA-100 gi%es a normal
result then with some e3ceptions primary haemostasis is intact (E3ceptions4 5P"$
Primary 5ecretion "efects$ mild Type 1 67". and so may ob%iate further screening of
platelet function#
J# f the PFA-100 is abnormal then formal platelet aggregation testing and platelet
nucleotide assays may be re@uired to establish the underlying cause#
A ne) auto*ated techni+ue for "latelet aggregation *easure*ent,
Abstract
A -ultistat Centrifugal Analyser (-CA! was used to measure platelet aggregation in
%itro# t has a capacity of about 90 samples per h# n the analyser platelet-rich plasma
and collagen-reagent were mi3ed$ and the turbidity was measured as a function of time#
The results were presented in arbitrary units ( arb # units!$ i#e# change in turbidity per
min > 1000# The estimated 0#:B reference range was B0-:B arb # units$ (n L 9C! and the
coeMcient of correlation between -CA results and results obtained by con%entional
aggregometry ( Fibromat ! was 0#)B (P less than 0#001!# The -CA method registered B0-
IBD reduction of platelet aggregation after inta,e of low dose acetylsalicylic acid (A5A!
(1#J-9#0 mg;,g! during K days in 1: subNects# The -CA method is suitable to monitor
A5A treatment routinely in order to establish an indi%idual appropriate A5A dose during
prophylactic treatment of arterial thromboembolic disease#
-"urious auto*ated "latelet count, nu*eration of .east for*s as "latelets
b. the cell-$/N 0000,
Abstract
7e recently encountered a patient with thrombocytopenia secondary to multiple drug
therapy$ disseminated prostatic adenocarcinoma$ and sepsis who had a sudden
decrease in his platelet count as enumerated by the Cell-"G? 9000 hematology analyEer
(Abbott "iagnostics$ 5anta Clara$ CA!# A manual platelet count performed thereafter was
e%en lower# The etiology of the spurious platelet count was clari0ed when numerous
yeast forms were obser%ed on routine microscopy of the peripheral blood smear#
5ubse@uently$ these organisms were identi0ed as Candida glabrata from a positi%e
blood culture (<ACTEC :J90$ <ecton "ic,inson$ Coc,eys%ille$ -"!# To our ,nowledge$
this is the 0rst report of spurious enumeration of yeast forms as platelets in an
automated hematology system# The principle underlying platelet enumeration by the
Cell-"G? 9000 system and other hematology analyEers and the %alue of microscopy on
peripheral smears with une3pected C<C count results are discussed#