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Flow cytometric analysis of thiazole orange uptake by platelets: a
diagnostic aid in the evaluation of thrombocytopenic disorders
J Kienast and G Schmitz
Department of Internal Medicine, University of Munster, FRG.
Thiazole orange (TO), a fluorescent dye originally synthesized for
reticulocyte analysis, is characterized by a large fluorescence enhancement
and high quantum yield on binding to nucleic acids, particularly RNA. In
addition, the dye readily permeates live cell membranes. We applied TO
staining, followed by fluorescence-activated flow cytometric analysis, to
platelets in whole blood samples from hematologically normal subjects and
patients with various quantitative platelet disorders. The percentage of
TO-positive platelets in 50 control subjects was 8.6 +/- 2.8% (mean +/- SD)
ranging from 2.8% to 15.8%. In 21 thrombocytopenic patients whose bone
marrow contained normal to increased numbers of megakaryocytes, the
percentage of fluorescently labeled platelets was significantly elevated (P
less than .0001) to 26.9 +/- 10.9% (range, 13.3% to 57.1%). In contrast,
the proportion of positively stained platelets in 23 patients with
thrombocytopenia due to impaired platelet production (various conditions
with reduced marrow megakaryocytes) did not significantly differ from the
controls, whereas the absolute counts of TO-positive platelets were
significantly lowered (P less than .0001). Differences in the distributions
of the percentage values as well as of the absolute counts for TO-positive
platelets between the two patient groups were again highly significant (P
less than .0001). Both the sensitivity and the specificity of this method
in distinguishing between these categories of thrombocytopenia were greater
than or equal to 95%. We conclude that flow cytometric analysis of
platelets after staining with TO is a sensitive and specific test that
rapidly provides information on the thrombopoietic activity in
thrombocytopenic disorders. Our data further suggest that increased amounts
of residual RNA characterize platelets released under conditions of "stress
thrombopoiesis."
Volume 75,
Issue 1,
pp. 116-121,
01/01/1990
Copyright © 1990 by The American Society of Hematology
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