Abstract
To establish a new method for the detection of platelet derived microparticles (PMPs) by cytometric bead array and to evaluate the characteristic of this method. Using detection beads coated with monoclonal antibody against platelet glycoprotein IIIa to capture PMPs, then using CD41 PE to react with PMPs captured by detection beads, finally using flow cytometer detect beads-PMPs-CD41 PE complex. Anticoagulant, reaction time and other condition were optimized. Then reproducibility and linear range of this method were evaluated. PMPs of 45 health adults were determined by this method. Detection beads were stable in 4 degrees C for over 90 days (coefficient of variance was 0.65%) which matched the basic request of clinical detection. CTAD could restrain the in vitro release of PMPs efficiently; The reaction could reach maximum combination at room temperature after 4 hour; Non-specific absorbance of detection beads could be eliminated by washing of PBS. This method had preferable reproducibility (CV in batch was 6.8%; CV between batch was 10.4%) and a wide linear range from 0 to 200 microl of platelet activation suspension (r = 0.9962). PMP level of 45 health adults was 1.031-1.766. PMPs can be detected exactly by cytometric bead array, and it's important for PMPs detection to be an assistant for diagnosis of haemorrhagic and thrombotic diseases.
Published Version
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