DNA微阵列芯片法与直接测序法检测CYP2C19基因型的比较研究

Comparison Analysis of CYP2C19 Polymorphisms Detection by DNA Microarray and DNA Microarray

目的用DNA微阵列芯片法和直接测序法两种方法检测氯吡格雷相关基因CYP2C19的突变情况，并进行比较分析。同时将基因型检测结果与临床资料进行分析，初步探讨CYP2C19基因型检测在氯吡格雷用药治疗中的临床意义。方法收集180例诊断为急性冠脉综合症并首次接受经皮冠状动脉介入治疗术（PCI）的全血标本。其中90例患者术后服用氯吡格雷之前，采用DNA微阵列芯片法和DNA直接测序法检测CYP2C19突变位点，确定其基因型。另外90例对照组患者使用氯吡格雷药物但不检测CYP2C19基因型。分析两组患者随访过程中发生冠脉血栓事件的差异。结果①两种检测方法准确度比较：在试验组90例患者中，DNA微阵列芯片法和DNA测序法均检出4种基因型组合：＊1／＊1（636GG，681GG），＊1／＊2（636GG，681GA），＊2／＊2（636GG，681AA）和＊1／＊3（636GA，681GG），分布频率分别为44例（48．9％），36例（40％），5例（5．6％）和5例（5．6％），而＊3／＊3（636AA，681GG）和＊2／＊3（636GA，681GA）未检测到。两种方法的准确度完全一致。②将检测结果与临床资料进行相关性分析：经CYP2C19基因型指导氯吡格雷用药的患者发生支架冠脉血栓事件的比例（0％）明显低于对照组（3．33％，P〈0．05）。结论①DNA微阵列芯片法灵敏度和准确度与DNA直接测序法（金标准方法）检测相符率为100％。②DNA微阵列芯片法是一种快速、准确发现CYP2C19突变位点，鉴别cYP2C19基因型的检测方法。③检测结果与临床相关性分析结果显示基因型指导氯吡格雷用药有助于患者用药剂量的调整，或选用其它抗凝血药物，从而可降低冠脉血栓事件的发生率。

Objective To compare and analyze the clinical application value of detection of Cytochrome P450 2C19 （CYP2C19） polymorphisms for clopidogrel application by DNA sequencing and DNA microarray. Methods 90 blood samples were randomly collected from patients,who were diagnosed as acute coronary syndrome and treated with percutaneous coronary intervention （PCI） for the first time. The genotyping of CYP2C19 ＊ 2 and CYP2C19 ＊ 3 alleles were performed in 90 samples through DNA sequencing and DNA microarray before using clopidogrel. The control group contains 90 patients with non-genotype guided clopidogrel usage. The clinical significance of detection of these polymorphisms was evaluated by comparing the proportion of patients, who suffered with the stent thrombosis between control and genotype-guided groups u- sing Chi square test. Results Both DNA sequencing and DNA microarray results showed that 4 kinds of gene type were detected： ＊1/＊1（636GG,681GG）, ＊ 1/＊ 2（636GG,681GA）, ＊2/＊ 2（636GG,681AA） and ＊ 1/＊ 3（636GA,681GG）,respectively. The distribution of them were 48.9% ,40% ,5.6% and 5.6% respectively. However, ＊ 3/＊ 3（636AA,681GG） and ＊ 2/＊ 3（636GA,681GA） were not found. Besides,in the studied group,the rate of stent thrombosis （0.0%） was significantly lower than that in the control group （3.3 %）. Conclusion （1）As to the sensibility and accuracy of CYP2C19 polymorphism detection,DNA sequencing coincides with DNA microarray. （2）The detection of CYP2C19 polymorphism by DNA microarray was validated to be rapid and reliable. （3）The genotyping of CYP2C19 with this method can effectively guide clo pidogrel application,and thus reduce the occurrence of stent thrombosis.