Abstract
BackgroundPrevious studies have shown that flow cytometry is a reliable test to quantify platelet function in stored platelet concentrates (PC). It is thought that flow cytometry is laborious and hence expensive. We have optimized the flow cytometry-based quantification of agonist induced platelet activation (PACT) to a labor, time and more cost-efficient test. Currently the quality of PCs is only monitored by visual inspection, because available assays are unreliable or too laborious for use in a clinical transfusion laboratory. Therefore, the PACT was applied to monitor PC activation during storage.Study design and methodsThe optimized PACT was used to monitor 5 PCs during 10 days of storage. In brief, optimized PACT uses a ready-to-use reaction mix, which is stable at -20°C. When needed, a test strip is thawed and platelet activation is initiated by mixing PC with PACT. PACT was based on the following agonists: adenosine diphosphate (ADP), collagen-related peptide (CRP) and thrombin receptor-activating peptide (TRAP-6). Platelet activation was measured as P-selectin expression. Light transmission aggregometry (LTA) was performed as a reference.ResultsBoth PACT and LTA showed platelet function decline during 10-day storage after stimulation with ADP and collagen/CRP; furthermore, PACT showed decreasing TRAP-induced activation. Major differences between the two tests are that PACT is able to measure the status of platelets in the absence of agonists, and it can differentiate between the number of activated platelets and the amount of activation, whereas LTA only measures aggregation in response to an agonist. Also, PACT is more time-efficient compared to LTA and allows high-throughput analysis.ConclusionPACT is an optimized platelet function test that can be used to monitor the activation of PCs. PACT has the same accuracy as LTA with regard to monitoring PCs, but it is superior to both LTA and conventional flow cytometry based tests with regard to labor-, time- and cost efficiency.
Highlights
The gold standard for testing platelet function is light transmission aggregometry (LTA) [1], which was described for the first time by Born and Cross in 1963 [2]
Major differences between the two tests are that platelet activation (PACT) is able to measure the status of platelets in the absence of agonists, and it can differentiate between the number of activated platelets and the amount of activation, whereas LTA only measures aggregation in PLOS ONE | DOI:10.1371/journal.pone
PACT is an optimized platelet function test that can be used to monitor the activation of platelet concentrates (PC)
Summary
The gold standard for testing platelet function is light transmission aggregometry (LTA) [1], which was described for the first time by Born and Cross in 1963 [2]. Other fast tests available in clinical labs, such as Multiplate (multiple electrode impedance aggregometry), rotational thromboelastometry (ROTEM), thromboelastography (TEG), the platelet function analyzer (PFA)-100 or the verifyNOW, are all based on the formation of a platelet aggregate or blood clot. These methods are not as laborious as LTA, but are unable to show pre-activation of platelets, do not allow high-throughput analysis and are insensitive in case of low platelet numbers [4]. The PACT was applied to monitor PC activation during storage
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