Balkan Journal of Medical Genetics

CO-EXISTENCE OF CYP2C19*1/*2 AND ABCB1C.3435 CT GENOTYPE HAS A POTENTIAL IMPACT ON CLINICAL OUTCOME IN CAD PATIENTS TREATED WITH CLOPIDOGREL
Nestorovska KA, Naumovska Z, Staninova Stojovska M, Sterjev Z, Dimovski A, Suturkova Lj
*Corresponding Author: Aleksandra Kapedanovska Nestorovska, PhD, Faculty of Pharmacy, Ss. Cyril and Methodius University in Skopje, Skopje, RN Macedonia, Mother Theresa str 47, 1000 Skopje, R. North Macedonia, Email address: alka@ff.ukim.edu.mk
page: 35

MATERIALS AND METHODS

Study population A total of 96 patients were included the study. Sam- ples from all patients were derived from the Special Hos- pital for Surgical Diseases “Filip II” in Skopje, R.N. Mac- edonia. The demographic and clinical characteristics of the patients enrolled in the study are presented in Table 1. Clopidogrel was administrated to all patients by the follow- ing regimen: a loading dose of 600 mg, on the first day of the treatment, and maintenance dose of 75 mg daily for up to 15 months. The follow up period was > 12 months. The primary endpoint of this study was the occurrence of the first clinical sign of the following MACE. The study was conducted in accordance with the Declaration of Helsinki and approved by the Ethics committee of the Faculty of Pharmacy- Skopje. DNA isolation Genomic DNA was isolated from leucocytes ob- tained (PBMCs) from 3ml peripheral blood collected by venepunction in vacutainers containing EDTA (Ethylen- ediaminetetraacetic acid) as an anticoagulant. DNA isola- tion was performed using the standard phenol/chloroform extraction protocol. Subsequently, the concentration of the obtained DNA was measured with the NanoDrop 2000c UV – Vis spectrophotometer (ThermoFisher Scientific, Wyman Street Waltham, MA USA). DNA purity was veri- fied by UV absorption at 260/280 nm, while DNA integrity was assessed using 1% agarose gel containing ethidium bromide. DNA samples were stored at 4°C. Genotyping The genotyping analysis for both polymorphisms (CYP2C19*2 and ABCB1 C3435T) was done by Real- Time PCR using the allelic discrimination method on the MxPro 3005P instrument (Agilent technologies, Santa Clara, CA, USA). The amplification and the detection of the specific SNPs alleles were performed using specific TaqMan Drug Metabolism Genotyping Assays according to manufacture recommendations (ThermoFisher Scien- tific, Foster City, CA, USA). The presence of the SNPs was determined using the MxPro Software v.5.1. Statistical analysis The obtained data was analyzed using SPSS software. Genotype distribution for the studied polymorphisms was in correlation with the Hardy-Weinberg equilibrium, ac- cording to the X 2 test. X 2 and Fischer exact probability test were used to compare the genotype distributions and allelic frequencies between the patient population and positive/ negative outcome. Odds ratios (OR) were calculated with 95% confidence interval limit (95% CI). P value ≤0.05 was considered as statistically significant.

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