焦磷酸测序技术对阿司匹林个体化用药相关SNP位点分型

发布时间：2018-05-22 11:35

本文选题：阿司匹林 + 焦磷酸测序技术　；参考：《中国药科大学学报》2017年05期

【摘要】：阿司匹林的抗血小板作用在临床存在显著的个体差异,而rs5918、rs12041331和rs730012 3个单核苷酸多态性位点与阿司匹林的药效和不良反应有着密切的联系。本文基于PCR结合焦磷酸测序技术,建立了阿司匹林相关SNP位点的基因分型方法。自主设计扩增引物和测序引物;优化PCR扩增条件,使3个位点能够在相同的条件下扩增并通过梯度稀释基因组DNA检测方法的灵敏度;最后,分别基于本文建立的新方法和Sanger测序技术对20例临床样本进行3个位点的基因分型,验证该方法的准确性。结果显示,本文成功建立了阿司匹林个体化用药相关SNP位点的基因分型方法,检测下限低至0.4 ng基因组DNA,用两种方法对20例临床样本进行3个位点的基因分型,结果完全一致,说明该方法有较高的准确性。
[Abstract]:There were significant individual differences in the antiplatelet effects of aspirin, and three single nucleotide polymorphisms (SNPs) of rs5918, rs12041331 and rs730012 were closely related to the pharmacodynamics and adverse reactions of aspirin. Based on PCR and pyrosequencing techniques, a genotyping method for aspirin associated SNP loci was established. The amplification primers and sequencing primers were designed independently; the PCR amplification conditions were optimized so that the three loci could be amplified under the same conditions and the sensitivity of the method was determined by gradient dilution of genomic DNA. Based on the new method established in this paper and Sanger sequencing technique, the three loci of 20 clinical samples were genotyped to verify the accuracy of the method. The results showed that the genotyping method of SNP loci associated with individual aspirin use was successfully established. The detection limit was as low as 0.4 ng genomic DNA.Twenty clinical samples were genotyped with three loci by two methods, and the results were consistent with each other. It shows that this method has high accuracy.
【作者单位】：中国药科大学药物质量与安全预警教育部重点实验室;河北省人民医院药学部;南京军区南京总医院药理科;
【基金】：国家自然科学基金资助项目(No.81673390) 江苏省重点研发计划资助项目(No.BE2016745) 江苏省自然科学基金资助项目(No.BK20151445) 药物质量与安全预警教育部重点实验室资助项目(No.DQCP2015MS02) 中国药科大学药学基地科研训练及科研能力提高资助项目(No.J1310032) 江苏省“青蓝工程”资助项目~~
【分类号】：R96

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