双色荧光杂交芯片技术检测CYP2C19单核苷酸多态性  

Application of dual-color fluorescence hybridization chip technique for single nucleotide polymorphism detection in CYP2C19

作　　者：邵华[1,2] 刘乃丰[3,4] 周东蕊[5] 程璐[6] 翟紫红[2] 蔡琳玲[2] 

机构地区：[1]中国药科大学药理研究室,江苏南京210009 [2]东南大学附属中大医院临床药理,江苏南京210009 [3]东南大学附属中大医院心脏科,江苏南京210009 [4]东南大学心血管病研究所,江苏南京210096 [5]东南大学儿童发展与学习科学教育部重点实验室,江苏南京210096 [6]东南大学生物电子学国家重点实验室,江苏南京210096

出　　处：《中国新药与临床杂志》2009年第4期249-253,共5页Chinese Journal of New Drugs and Clinical Remedies

基　　金：江苏省卫生厅奥赛康基金项目(P200603)

摘　　要：目的应用双色荧光杂交芯片技术检测CYP2C19单核苷酸多态性,为相关研究提供方法和依据。方法设计丙烯酰胺标记引物,以87位中国江苏地区汉族健康志愿者的基因组DNA为模板进行目标序列的PCR扩增,并将产物固定于氨基玻片上,变性后再与荧光标记的寡核苷酸探针进行杂交,通过基因芯片扫描仪检测荧光信号,判断CYP2C19基因型,并采用DNA直接测序法验证结果。结果所有芯片法检测结果均经DNA直接测序验证,2种方法分型结果完全一致。87位志愿者中CYP2C19*2和CYP2C19*3的发生频率分别为9%和42%,其中CYP2C19弱代谢者(CYP2C19*2/*3、*3/*3)的发生频率为8%。结论双色荧光杂交芯片技术检测CYP2C19单核苷酸多态性特异性高,其高通量特点适用于大规模人群的检测。AIM To detect CYP2C19 single nucleotide polymorphisms （SNP） by dual-color fluorescence hybridization chip technique. METHODS Acrylamide-modified primers were designed and genomes from 87 healthy Han Chinese volunteers of Jiangsu area were used as PCR templates. After attached to the acrylmodified slide surface, PCR products were denatured and rehybridized to the oligonucleotide fluorescent probes labeled with Cy3 and Cy5 dyes. The CYP2C19 genotypes were determined based on the fluorescent signals of microarray scanner and direct DNA sequencing was performed to evaluate the genotyping validity. RESULTS The genotyping results of dual-color fluorescence hybridization chip on 87 samples were completely consistent with the results of direct DNA sequencing. The allelic frequencies of CYP2C19^＊2 and CYP2C19^＊3 were 9% and 42%, respectively, and the frequency of the poor metabolizer of CYP2C19 （CYP2C19^＊2/^＊3,^＊3/^＊3） was 8%. CONCLUSION Dual-color fluorescence hybridization chip technique is a specific method for SNP detection in CYP2C19, the high-throughput characteristic make it a favorable method for SNP genotyping in a large population.

关 键 词：原位杂交 荧光 芯片分析技术 多态性 单核苷酸 CYP2C19 

分 类 号：R394.6[医药卫生—医学遗传学]