A Simple Test for Measuring Platelet Aggregation

Summary

A simple method of measuring platelet aggregation is described which is suitable as a screening test in studies of haemostasis. This test is based on the phenomenon that blood platelets present in blood in the form of aggregates, are more rapidly deposited during centrifugation than platelets present in dispersed form.

A comparative study was made of platelet concentration in the supernatant plasma of slowly centrifuged EDTA-blood, heparinized blood and heparinized blood mixed with ADP (end concentration 4.5 μg/ml).

As expected, the platelet concentration was always highest in the EDTA-plasma because EDTA prevents platelet aggregation and disperses platelet aggregates if already present. The difference in platelet count between EDTA-plasma and heparin-ADP, expressed as per cents of the count in EDTA-plasma, was called ADP-aggregation. The ADP-aggregation exceeded 79% in 35 normal subjects. It was lower than 79% in 6 of 9 von Willebrand patients, 11 of 13 relatives of these patients and 7 of 8 uraemic patients.

The percentual difference in platelet concentration between EDTA-plasma and heparinized plasma was called spontaneous aggregation. This was significantly lower in von Willebrand patients and their relatives than in the control group. By comparing all groups of patients collectively with the normal control group, an even more marked difference in spontaneous aggregation was found.

• References

• 1 Bagnold R. A. Experiments on a gravity-free dispersion of large solid spheres in a Newtonian fluid under shear. Proc. roy. Soc. Lond. A 225: 49 1954;
• 2 Borchgrevink C. F. A method for measuring platelet adhesiveness in vivo . Acta med. scand. 168: 157 1960;
• 3 Born G. V. R. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature (Lond.) 194: 927 1962;
• 4 Born G. V. R, Gross M. J. The aggregation of blood platelets. J. Physiol. 168: 178 1963;
• 5 Bull B. S. M, Zucker B. Changes in platelet volume produced by temperature, metabolic inhibitors and aggregating agents. Proc. Soc. exp. Biol. (N. Y.) 120: 296 1965;
• 6 Gaspary E. A. Effect of red blood-cells and adenosine on platelet adhesiveness. Lancet II: 1273 1965;
• 7 Castaldi P. A, Rozenberg M. C, Stewart J. H. The bleeding disorder of uraemia. Lancet II: 66 1966;
• 8 Constantine J. W. Temperature and the aggregation of rabbit platelets by adenosine diphosphate. Nature (Lond.) 205: 1075 1965;
• 9 Creveld S. van, Pascha C. N. Ziekte van von Willebrand en enige andere toestanden gecombi-neerd met een “nieuwe” afwijking in de functie der bloedplaatjes. Ned. T. Geneesk. 112: 646 1968;
• 10 Cronberg S, Nilsson I. M, Silver J. Studies on the platelet adhesiveness in von Willebrand’s disease. Acta med. scand. 180: 43 1966;
• 11 Davey M. G, Lüscher E. F. Biochemical aspects of platelet function and hemostasis. Seminars in Hematology 5: 5 1968;
• 12 Gaarder A, Jonsen J, Laland S, Hellem A, Owren P. A. Adenosine diphosphate in red cells as a factor in the adhesiveness of human blood platelets. Nature (Lond.) 192: 531 1961;
• 13 Haanen C, Holdrinet A. Blood platelet aggregation under various conditions and in some diseases. Exp. Biology and Med. Vol. 3: 164 Karger, Basel; New York: 1968
• 14 Hardisty R. M, Hutton R. A. Bleeding tendency associated with “new” abnormality of platelet behaviour. Lancet I: 983 1967;
• 15 Harrison M. J. G, Mitchell J. R. A. The influence of red blood-cells on platelet adhesiveness. Lancet II: 1163 1966;
• 16 Hellem A. J, Ödegaard A. E, Skålhegg B. A. Investigations on adenosine diphosphate (ADP) induced platelet adhesiveness in vitro. I. The ADP-platelet reaction in various experimental conditions. Thrombosis et Diathesis Haemorrhagiga 10: 61 1963;
• 17 Hellem A. J, Ödegaard A. E, Skålhegg B. A. Platelet adhesiveness in chronic renal failure. Abstract: X^th Congress Internat. Soc. Haematol., Stockholm Pg. K: 1 1964;
• 18 Hjort P. F, Rapaport S. I, Owren P.A. A simple, specific one-stage prothrombin assay using Russell’s viper venom in cephalin suspension. J. Lab. clin. Med. 46: 89 1955;
• 19 Holdrinet A, Ewals M, Haanen G. Increase in platelet aggregation with time after venepuncture. Lancet II: 893 1967;
• 20 Holdrinet A. Unpublished observation.
• 21 Holdrinet A, Ewals M, Haanen G. Platelet function and von Willebrand’s disease. Lancet I: 1150 1968;
• 22 Larrieu M. J, Caen J. P, Meyer D. O, Vainer H, Sidtan Y, Bernards J. Congenital bleeding disorders with long bleeding time and normal platelet count : II von Willebrand’s disease (Report of thirty-seven patients). Amer. J. Med. 45: 354 1968;
• 23 Mannucci P. M, Sharp A.A. Platelet volume and shape in relation to aggregation and adhesion. Brit. J. Haemat. 13: 604 1967;
• 24 Mattern G. F, Brackett F. S, Olson B. J. Determination of number and size of particles by electronic gating: blood cells. J. appi. Physiol. 10: 56 1957;
• 25 Niemeyer G, Reuter H, Gross R. Untersuchungen über die Aggregation menschlicher Blutplättchen. II. über den Einfluß der Temperatur auf die Plättchenaggregation. Klin. Wschr. 22: 1142 1967;
• 26 O’Brien J. R. Platelet aggregation, Part II, Some results from a new method of study. J. clin. Path. 25: 452 1962;
• 27 O’Brien J. R. Platelets : A Portsmouth syndrome?. Lancet II: 258 1967;
• 28 O’Brien J. R, Heywood J. B. Some interactions between human platelets and glass: von Willebrand’s disease compared with normal. J. clin. Path. 20: 56 1967;
• 29 Pruden E. L, Winstead M. E. Accuracy control of blood cell counts with the Coulter counter. Amer. J. med. Technol. 30: 1 1964;
• 30 Rozenberg M. G, Firkin B. G. The rate of platelet aggregation in haemorrhagic disease and thrombosis. Scand. J. Haemat. 3: 5 1966;
• 31 Salzman E. W. Measurement of platelet adhesiveness. A simple in vitro technique demonstrating abnormality in von Willebrand’s disease. J. Lab. clin. Med. 62: 724 1963;
• 32 Salzman E. W, Néri L. L. Adhesiviness of blood platelets in uremia. Thrombosis et Diathesis Haemorrhagiga 15: 84 1966;
• 33 Stewart J. H, Gastaldi P. A. Uraemic bleeding: a reversible defect corrected by dialysis. Quart. J. Med. 36: 409 1967;
• 34 Stormorken H. The defect in von Willebrand’s disease. In: Physiology of haemostasis and thrombosis. Conference held in conjunction with the 14^th annual symposium on blood. 179 Charles C. Thomas; Springfield, Ill. USA.: 1967
• 35 Strauss H. S, Bloom G. E. Von Willebrand’s disease. Use of a platelet adhesiveness test in diagnosis and family investigation. New Engl. J. Med. 273: 171 1965;
• 36 Vainer H, Caen J. P. Utilisation d’un test photométrique pour l’étude de l’effet de l’ ADP sur les plaquettes sanguines. Nouv. Rev. franc. Hémat. 3: 149 1963;
• 37 de Vries S. I, ten Cate J. W, den Hartog-Veerman M. E, van Dooren M. M. Quelques remarques sur la thrombocytopathie acquise chez les malades urémiques. Schweiz, med. Wschr. 98: 1689 1968;
• 38 Weiss H. J. Platelet aggregation, adhesion and adenosine diphosphate release in thrombo-pathia (platelet factor 3 deficiency). Amer. J. Med. 43: 570 1967;
• 39 Zucker M. B. In vitro abnormality of the blood in von Willebrand’s disease correctable by normal plasma. Nature (Lond.) 197: 60 1963;