焦磷酸测序检测CYP2C19基因多态性方法的建立

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Abstract [Abstract] Objective To establish a fast detection method for CYP2C19 genotyping based on pyrosequencing technology. Methods Genomic DNA was extracted from 50 blood samples of healthy subjects collected from December 2016 to March 2017 in Guizhou Provincial People′s Hospital, and a set of amplification primers and pyrophosphate sequencing primers were designed according to the single nucleotide polymorphism (SNP) sites of CYP2C19*2 (681G>A), CYP2C19*3 (636G>A), CYP2C19*17 (-806C>T). Single-stranded templates were made from PCR products, then sequencing and genotyping analysis were performed on a Qiagen PyroMark Q24 Sequencer System. The sequencing results were verified via Sanger sequencing. Results Based on the type and peak height of bases of DNA sequence, the genotypes of three SNPs could be identified clearly, which were consistent with Sanger sequencing results. Among the 50 samples of healthy group, 1 case was ultra-rapidmetabolism, 22 cases were rapidmetabolism, 22 cases were medium metabolism and 5 cases were slow metabolism. Conclusion The detection method of CYP2C19 polymorphism based on pyrosequencing technology is quick, accurate and convenient, which can be used to analyze the genotyping of CYP2C19 gene polymorphism.

Key words： CYP2C19 Gene polymorphism Pyrosequencing Clopidogrel

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	YANG Nannan1 XU Qin1 LUO Zhenyuan2 FU Xue2 HUANG Shengwen2▲

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YANG Nannan1 XU Qin1 LUO Zhenyuan2 FU Xue2 HUANG Shengwen2▲. Establishment of genotyping methods for polymorphisms of CYP2C19 gene based on pyrosequencing technology[J]. 中国医药导报, 2017, 14(29): 11-14.

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http://www.yiyaodaobao.com.cn/EN/ OR http://www.yiyaodaobao.com.cn/EN/Y2017/V14/I29/11

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