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Background: A reliable laboratory test to monitor on-clopidogrel platelet reactivity (PR) is very necessary. In addition, genetic factors also play an important part in on-clopidogrel PR. This study aimed to modify the original impedance whole blood platelet aggregation assay associated with the release assay to monitor on-clopidogrel PR and assess their relationship with genotype.
Methods: We adjusted the concentration of calcium in the in vitro reaction system of platelet aggregation to modify the original impedance whole blood platelet aggregation assay. Meanwhile, chronolume, which quantified the adenosine triphosphate (ATP) released from platelet dense granules, is added to this reaction system to reflect the platelet release function. In the modified assay, platelet magnified activation time (MAT) and the maximal platelet ATP release value (RV) were used to reflect platelet function parameters. In the original assay, the electrical resistance (Ω) and RV were used to reflect platelet function parameters. On-clopidogrel PR was detected by the original impedance whole blood platelet aggregation assay, modified assay, and flow cytometric vasodilator stimulated phosphoprotein (VASP) assay in 168 patients with acute coronary syndromes (ACS). CYP2C19*2 and CYP2C19*3 polymorphisms were also detected in all of these patients.
Results: This modified method showed that when 12.5μL CaCl2 (0.2 mmol/L) was added to the reaction system, MAT was appropriate (93 ± 23 seconds). The CVs for the modified impedance assay and release assay were 9.31% and 6.13%, respectively. The mean VASP-PRI in the patient group treated with clopidogrel was significantly lower than that in the control group without antiplatelet therapy (54.88 ± 16.81% vs. 79.86 ± 10.24%, p < 0.001). MAT of the modified method in VASP PRI > 50% group were shorter than that in the PRI < 50% group [185 (154 - 241) vs. 214 (184 - 250), p < 0.05]. Meanwhile, the RV of the modified method in VASP PRI > 50% group were higher than that in the PRI < 50% group [1.00 (0.72 - 1.47) vs. 0.82 (0.62 - 1.08), p < 0.05]. However, the electrical resistance (Ω) and RV of the original method showed no differences between the two groups [0 (0 - 2) vs. 0 (0 - 1.25), 0.05 (0 - 0.25) vs. 0.08 (0 - 0.24); p > 0.05, p > 0.05, respectively). Moreover, neither the original method nor the modified method showed differences in patients with CYP2C19 (*2 and *3) wild type and mutant type.
Conclusions: The consistency of the modified assay and VASP assay is good. The modified assay may be a potentially good laboratory method to monitor antiplatelet therapy.
DOI: 10.7754/Clin.Lab.2015.150403
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