In vitro effect of blood cell counts on multiple-electrode impedance aggregometry in dogs

Platelet function testing in whole blood is widely used to evaluate the effect of antiplatelet agents, but it is not known whether results are affected by whole blood parameters. This study investigated the importance of platelet count, haematocrit, red blood cells (RBC), and white blood cells in whole blood platelet aggregometry. We included 417 patients with coronary artery disease on aspirin mono-therapy and 21 aspirin-naïve healthy individuals. Blood sampling was performed one hour after aspirin ingestion. The antiplatelet effect of aspirin was evaluated using the VerifyNow® Aspirin assay and multiple electrode aggregometry (MEA, Multiplate®) induced by collagen (1.0 μg/mL) and arachidonic acid (1.0 or 0.75 mmol/L). Measurements of whole blood parameters were performed to evaluate the three major cell lines in circulating blood. In patients, platelet count correlated significantly with platelet aggregation (MEA(collagen), p<0.0001; MEA(arachidonic acid), p<0.0001; VerifyNow®, p=0.03). Haematocrit and RBC correlated inversely with MEA induced by collagen (p(haematocrit)<0.001; p(RBC)=0.07) and with VerifyNow® (p(haematocrit)<0.0001; p(RBC)<0.0001), but not with MEA induced by arachidonic acid (p(haematocrit)=1; p(RBC)=0.87). White blood cells correlated significantly with platelet aggregation (MEA(collagen), p<0.001; MEA(arachidonic acid), p<0.0001; VerifyNow®, p=0.05). Similar associations were observed in aspirin-naïve healthy individuals. Whole blood aggregometry is dependent on all major cell lines in whole blood. Importantly, platelet aggregation is significantly associated with platelet count even within the normal range. Copyright © 2011 Elsevier Ltd. All rights reserved.

Introduction Sepsis leads to an activation of the immune system and hemostatis. However, studies on platelet aggregation in severe sepsis using impedance aggregometry have not been performed and the diagnostic and prognostic capabilities are unknown. In the present study we hypothesized that impedance aggregometry findings might serve as a biomarker for the diagnosis and prognosis of severe sepsis. Methods Eighty patients with severe sepsis and 50 postoperative patients were included in the prospective observational study. Platelet function was determined at the first day of severe sepsis and surgery, respectively, using impedance aggregometry (Multiplate®). Moreover, platelet count, procalcitonin, interleukin 6, C-reactive protein and 30-day mortality were determined. Results Compared to postoperative patients, platelet aggregation was significantly reduced in patients with severe sepsis (collagen-test: 70.8 (44.4, 83.2) arbitrary units (A.U.) vs. 26.8 (12.7, 45.8) A.U.; P <0.001; median and quartiles). Furthermore, marked differences in platelet function were observed in survivors and non-survivors of severe sepsis (collagen-test: 33.4 (10.9, 48.8) A.U. vs. 12.4 (6.5, 25.0) A.U.; P = 0.001). Kaplan-Meier analysis demonstrated that higher platelet function was associated with a mortality of 10%, while mortality was 40% when platelet function was low (collagen-test; P = 0.002). The odds ratio was 6.0. In both univariate and multivariate analyses (including procalcitonin, IL6, C-reactive protein and platelet count) impedance aggregometry using collagen as the activator proved to be the best and an independent predictor for the diagnosis and prognosis of severe sepsis in critical illness. Conclusions In severe sepsis, impedance aggregometry allows better prediction of diagnosis and survival than conventional biomarkers and platelet count.