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Light Transmission Aggregometry (LTA) on TA-8V (Stago®): What about Normal Values?

A. Dericquebourg1, M. Daniel2, J.-C. Bordet1, F. Sobas1, C. Nougier1, C. Negrier1,2, S. Le Quellec1,2

1Service d’Hématologie Biologique, Centre de Biologie et Pathologie Est, Hospices Civils de Lyon, Lyon, France, 2Unité d’Hémostase Clinique, Hôpital Cardiologique Louis Pradel, Hospices Civils de Lyon, Lyon, France

Abstract Number: PB0895

Meeting: ISTH 2021 Congress

Theme: Platelet Disorders, von Willebrand Disease and Thrombotic Microangiopathies » Platelet Function Disorders, Hereditary

Background: Diagnosis of platelet function disorders relies on light-transmission aggregometry (LTA) using a panel of agonists. Hence, assessment of normal values and laboratory performances of the reagents used are critical for clinical practice.

Aims: The aim was to evaluate the performances of collagen, ADP, arachidonic acid,  TRAP-6, and ristocetin reagents (Stago®, Asnières-sur-Seine, France) using LTA method in our laboratory.

Methods: LTA was performed on TA-8V aggregometer (SD-Medical, Stago®) on citrated platelet-rich plasma (PRP) obtained from 30 consenting healthy volunteers (NCT04269603), using 5 different agonists at several final concentrations: 2µg/mL collagen, 2 and 5µM ADP , 1mM arachidonic acid, 10µM TRAP-6 and 1.2 mg/mL ristocetin.
Normal values, intra-assay variability (CVintra, %) and inter-operator variability for maximal aggregation intensity (Imax, %) were measured after platelet activation by all agonists. Reagent stability was tested over a period of 2 months after reconstitution, sampling, and storage prior to use.

Results: Normal values for Imax ranged from >30% for 2µM ADP- to >80% for arachidonic acid- and ristocetin-induced platelet activation. Intra-assay variability was good, being below 8% in all conditions for platelet activation (range 2.7 – 7.8%). These results were in line with manufacturer’s normal values (when available; table1).
Inter-operator variability showed an agreement of >80% (range 84.6 – 100%) for all agonists tested, with regards to Imax values, and reached 100% when aggregation curves were examined by a blinded expert biologist. All reagents were stable for 2 months.

Agonist Imax normal values, % CVintra, % Stability after reconstitution and sampling, months (storage temperature)
Manufacturer In house Manufacturer  In house Manufacturer  In house
Collagen 2 µg/mL >66 >73 5.8 2.7 ND 2 (2-8°C)
ADP 2 µM >47 >30 6.4 7.8 ND 2 (-20°C)
ADP 5 µM ND >65 ND 3.1 ND 2 (-20°C)
Arachidonic acid 1 mM >68 >80 3.8 4.2 ND 2 (-20°C)
TRAP-6 10 µM >69 >66 4.2 5.7 ND 2 (-20°C)
Ristocetin 1.2 mg/ml ND >80 ND 2.5 ND 2 (-20°C)

Maximum aggregation intensity (Imax) normal values and intra-assay variability (CV) of various agonist-induced platelet activation using TA-8V light-transmission aggregometer

ND: non determined

Conclusions: We provided normal range values and a study design for the assessment of laboratory performance of LTA method using  collagen-, ADP-, arachidonic acid-, TRAP-6-, and ristocetin-agonists for platelet activation. The low CVintra, the good agreement between operators, and the 2-months period stability for all reagents tested, make them suitable for routine screening of platelet function disorders.

To cite this abstract in AMA style:

Dericquebourg A, Daniel M, Bordet J-, Sobas F, Nougier C, Negrier C, Le Quellec S. Light Transmission Aggregometry (LTA) on TA-8V (Stago®): What about Normal Values? [abstract]. Res Pract Thromb Haemost. 2021; 5 (Suppl 2). https://abstracts.isth.org/abstract/light-transmission-aggregometry-lta-on-ta-8v-stago-what-about-normal-values/. Accessed August 9, 2022.

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