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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。
Objective To detect the single nucleotide polymorphism of CYP2C19 by using two-color fluorescence hybridization chip technology, and to provide methods and evidences for related research. Methods Acrylamide-labeled primers were designed and used to amplify the target sequence of genomic DNA from 87 healthy volunteers in Han nationality of Jiangsu Province in China. The products were fixed on glass slides, then denatured and incubated with fluorescently labeled oligonucleotides The probes were hybridized and the fluorescence signal was detected by a gene chip scanner to determine the CYP2C19 genotype. The results were verified by DNA direct sequencing. Results All the chip test results were verified by DNA sequencing, the results of two methods were exactly the same. The frequency of occurrence of CYP2C19 * 2 and CYP2C19 * 3 in 87 volunteers was 9% and 42%, respectively. The frequency of CYP2C19 weak metabolizers (CYP2C19 * 2 / * 3, * 3 / * 3) was 8% in 87 volunteers. Conclusion The two-color fluorescence hybridization chip technique has high specificity for detecting single nucleotide polymorphism of CYP2C19, and its high-throughput characteristics are suitable for the detection of large-scale population.