Abstract: Objective: To investigate the preparation process of gene chips for individualized treatment of thrombotic diseases (TD),and to elucidate a rapid and high-throughput method for detecting the drug metabolism genes of clopidogrel and warfarin. Methods: Different kinds of probes and primers were designed according to the clopidogrel drug metabolism gene sites (CYP3A4,CYP2C19*17,CYP2C19*2, and CYP2C19*3) and warfarin drug metabolism gene sites (CYP4F2*3,GGCX,VKORC1-2,VKORC1-1,CYP2C9*2, and CYP2C9*3) to prepare the gene chip detection kit for individualized treatment of TD.The steps of microarray detection were DNA extraction,PCR amplification,hybridization,elution and scanning.The sensitivity,repeatability and stability of the gene chips were evaluated. The samples of 150 subjects with TD from three regions were selected,and the drug metabolism genes were detected.Based on the medical guidances in the clinic and the disease recovery status after 6 months,the effectiveness of gene chip detection kit was determined. Results: The effective hybridization signals were observed in the metabolism genes of two kinds of drugs detected by gene chips;the minimum detection concentration of PCR amplification products was 10³ copies·mL^-1 detected by gene chip;the coincidence rate of simultaneous determination of three chips in different batches and parallel measurement of 10 chips in the same batch was 100%;the chip was stable and could be stored for at least 6 months;the effective rate of gene chip detection kit in the 150 TD subjects from three regions was more than 90%. Conclusion: The high-throughput gene chip detection kit of individualized treatment of TD to detect the drug metabolism genes of clopidogrel and warfarin are successfully established,and the method has high sensitivity,satisfactory repeatability,strong stability and excellent preparation effect.

Key words: thrombotic disease, gene chip, clopidogrel, warfarin