Recommendations for the standardization of light transmission aggregometry: a consensus of the working party from the platelet physiology subcommittee of SSC/ISTH

Cattaneo, Marco; Cerletti, Chiara; Harrison, Paul; Hayward, Catherine P.M.; Kenny, Dermot; Nugent, Diane J.; Nurden, Paquita; Rao, A. Koneti; Schmaier, Alvin H.; Watson, Steve P.; Lussana, Federico; Pugliano, Mariateresa; Michelson, Alan D.

doi:10.1111/jth.12231

Cited by 447 publications

(495 citation statements)

References 31 publications

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“…As well as requiring expensive and dedicated aggregometers, the technique is labour-intensive and requires specialist training of the individual running the tests. Furthermore, though there are clear International Society on Thrombosis and Haemostasis (ISTH) guidelines for the use of various platelet agonists and their concentrations, adherence to these guidelines is poor and varies with local practice and regulations (6,7). Taking together these variables in assay methodology result in data that is very hard to compare and/or share between different sites and operators.…”

Section: The Principles Of Platelet Aggregometrymentioning

confidence: 99%

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96-well plate-based aggregometry

2018

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While there are many bench and bedside tests to assess platelet reactivity, ex vivo light transmission aggregometry (LTA) remains the gold standard. LTA, however, is expensive, time-consuming and requires dedicated equipment and staff, making it impractical in many situations. In addition, there is significant variability between data generated at different testing sites meaning that tests often need to be repeated if a patient is transferred to the care of a different hospital. As such, there is clearly an unmet need for standardization of platelet testing.Using the principles of LTA, aggregometry can be conducted in 96-well plates with readings being made in a standard plate reader. This approach allows for the assessment of multiple concentrations of agonists, since the volume of platelets required for each test is significantly lower than for LTA. Furthermore, the lyophilization of a set panel of agonists to a 96-well plate to produce a stable assay substrate allows the production of portable, standardized plates that can be used to generate reproducible tests at multiple sites.In this review, we will discuss the methods and uses of 96-well plate aggregometry for both research and the clinic.

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Section: The Principles Of Platelet Aggregometrymentioning

confidence: 99%

“…To perform the test, blood is taken into anti-coagulant and PRP and PPP isolated as per ISTH guidelines (6). About 40 μl of PRP or PPP is added to the appropriate wells of the Optimul plate and the plate is placed onto a BioShake IQ (QInstruments, Jena, Germany) thermomixer with 1200 rpm shaking at 37°C.…”

Section: Lyophilized Agonist Platelet Aggregationmentioning

confidence: 99%

96-well plate-based aggregometry

2018

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“…[10][11][12][13][14][15][16][17] Lighttransmission aggregometry has been widely used as the gold standard to identify and diagnose defects in platelet function. 12,13 However, it requires platelet-rich plasma, a time-consuming test procedure, and large sample volumes. Platelet aggregation can be measured in anticoagulated whole blood using electrical impedance.…”

Section: Introductionmentioning

confidence: 99%

Migration distance-based platelet function analysis in a microfluidic system

2013

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Aggregation and adhesion of platelets to the vascular wall are shear-dependent processes that play critical roles in hemostasis and thrombosis at vascular injury sites. In this study, we designed a simple and rapid assay of platelet aggregation and adhesion in a microfluidic system. A shearing mechanism using a rotating stirrer provided adjustable shear rate and shearing time and induced platelet activation. When sheared blood was driven through the microchannel under vacuum pressure, shear-activated platelets adhered to a collagen-coated surface, causing blood flow to significantly slow and eventually stop. To measure platelet adhesion and aggregation, the migration distance (MD) of blood through the microchannel was monitored. As the microstirrer speed increased, MD initially decreased exponentially but then increased beyond a critical rpm. For platelet-excluded blood samples, there were no changes in MD with increasing stirrer speed. These findings imply that the stirrer provided sufficiently high shear to activate platelets and that blood MD is a potentially valuable index for measuring the shear-dependence of platelet activation. Our microfluidic system is quick and simple, while providing a precise assay to measure the effects of shear on platelet aggregation and adhesion.

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“…16 A major problem that hindered progress in the field is that the definition itself of platelet dysfunction has remained vague. This has been because, despite recent and important efforts to reach a consensus among experts, 16,17 we do not have validated criteria for recognizing and classifying these conditions. Moreover, our ability to predict the risk of bleeding in the individual patient remains limited and, therefore, we are often in doubt as to whether or not to administer prophylactic treatments in situations of hemostatic challenges, such as giving birth or surgery.…”

mentioning

confidence: 99%