title: Comparison of Modified Impedance Whole Blood Platelet Aggregation Method Detecting Platelet Function in ACS Patients with Different CYP2C19 Genotypes.
abstract: BACKGROUND
  A reliable laboratory test to monitor onclopidogrel platelet reactivity (PR) is very necessary. In addition, genetic factors also play an important part in onclopidogrel PR. This study aimed to modify the original impedance whole blood platelet aggregation assay associated with the release assay to monitor onclopidogrel 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 (omega) and RV were used to reflect platelet function parameters. Onclopidogrel 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 12.5 microL 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 < 0.051. 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 < 0.051. However, the electrical resistance (omega) 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.
authors.forename: Chanjuan
authors.initials: C
authors.lastname: Cui
authors.forename: Rui
authors.initials: R
authors.lastname: Qiao
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
mesh_terms: D054058:Acute Coronary Syndrome / Q000097:blood / Q000175:diagnosis / Q000188:drug therapy*; D000255:Adenosine Triphosphate / Q000097:blood; D000328:Adult; D000368:Aged; D057205:Automation, Laboratory; D015415:Biomarkers / Q000097:blood; D001711:Biotransformation; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism; D002122:Calcium Chloride / Q000097:blood; D016022:Case-Control Studies; D015815:Cell Adhesion Molecules / Q000097:blood; D000077144:Clopidogrel; D065731:Cytochrome P-450 CYP2C19 / Q000235:genetics* / Q000378:metabolism; D016903:Drug Monitoring / Q000379:methods*; D017097:Electric Impedance; D005260:Female; D005838:Genotype; D006801:Humans; D008297:Male; D008840:Microfilament Proteins / Q000097:blood; D008875:Middle Aged; D010597:Pharmacogenetics; D010641:Phenotype; D010750:Phosphoproteins / Q000097:blood; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000493:pharmacokinetics / Q000627:therapeutic use*; D010979:Platelet Function Tests / Q000379:methods*; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D013988:Ticlopidine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000627:therapeutic use; D016896:Treatment Outcome; D055815:Young Adult; D000098662:Vasodilator-Stimulated Phosphoprotein
chemical_list: D015415:Biomarkers; D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; D010750:Phosphoproteins; D010975:Platelet Aggregation Inhibitors; D000098662:Vasodilator-Stimulated Phosphoprotein; D000255:Adenosine Triphosphate; D000077144:Clopidogrel; C045793:CYP2C19 protein, human; D065731:Cytochrome P-450 CYP2C19; D002122:Calcium Chloride; D013988:Ticlopidine