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Optical multichannel (optimul) platelet aggregometry in 96-well plates as an additional method of platelet reactivity testing.
- Source :
- Platelets; Nov2011, Vol. 22 Issue 7, p485-494, 10p, 3 Charts, 5 Graphs
- Publication Year :
- 2011
-
Abstract
- Platelet reactivity testing is important for the diagnosis of bleeding disorders, and increasingly to optimise anti-platelet therapy. Traditional light transmission aggregometry is considered the gold standard, whilst 96-well plate aggregometry, founded on similar principles, provides a higher throughput screening method. Despite the widespread use of both, methodologies and outputs vary widely between laboratories. We report a methodological approach towards providing a standardised optical detection of platelet aggregation (optimul method) based upon 96-well plate aggregometry. Individual wells of half-area 96-well plates were coated with gelatine and one of seven concentrations of arachidonic acid (AA), adenosine diphosphate (ADP), collagen, epinephrine (EPI), ristocetin, TRAP-6 amide or U46619, before being lyophilised, vacuum-sealed, foil-packed and stored at room temperature for up to 24 weeks. For platelet testing, 40 µl of platelet-rich plasma was added to each well. Platelet aggregation was determined by changes in light absorbance, release of ATP/ADP by luminescence and release of thromboxane (TX) A<subscript>2</subscript> by ELISA. Some experiments were conducted in the presence of aspirin (30 µM) or prasugrel active metabolite (PAM; 3 µM). Optimul plates stored for up to 12 weeks permitted reliable detection of concentration-dependent platelet aggregation, ATP/ADP release and TXA<subscript>2</subscript> production. PAM caused reductions in platelet responses to AA, ADP, collagen, EPI, TRAP-6 and U46619, whilst aspirin inhibited responses to AA, collagen and EPI. We conclude that the optimul method offers a viable, standardised approach, allowing platelet reactivity testing and could provide a broad platelet function analysis without the need for dedicated equipment. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 09537104
- Volume :
- 22
- Issue :
- 7
- Database :
- Complementary Index
- Journal :
- Platelets
- Publication Type :
- Academic Journal
- Accession number :
- 66310989
- Full Text :
- https://doi.org/10.3109/09537104.2011.592958