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焦磷酸测序技术对阿司匹林个体化用药相关SNP位点分型

邢晓清, 卜修民, 初亚男, 邹秉杰, 宋沁馨, 周国华

邢晓清, 卜修民, 初亚男, 邹秉杰, 宋沁馨, 周国华. 焦磷酸测序技术对阿司匹林个体化用药相关SNP位点分型[J]. 中国药科大学学报, 2017, 48(5): 577-582. DOI: 10.11665/j.issn.1000-5048.20170512
引用本文: 邢晓清, 卜修民, 初亚男, 邹秉杰, 宋沁馨, 周国华. 焦磷酸测序技术对阿司匹林个体化用药相关SNP位点分型[J]. 中国药科大学学报, 2017, 48(5): 577-582. DOI: 10.11665/j.issn.1000-5048.20170512
shu
XING Xiaoqing, BU Xiumin, CHU Yanan, ZOU Bingjie, SONG Qinxin, ZHOU Guohua. Genotype method for SNPs related to individualized aspirin treatment based on pyrosequencing technology[J]. Journal of China Pharmaceutical University, 2017, 48(5): 577-582. DOI: 10.11665/j.issn.1000-5048.20170512
Citation: XING Xiaoqing, BU Xiumin, CHU Yanan, ZOU Bingjie, SONG Qinxin, ZHOU Guohua. Genotype method for SNPs related to individualized aspirin treatment based on pyrosequencing technology[J]. Journal of China Pharmaceutical University, 2017, 48(5): 577-582. DOI: 10.11665/j.issn.1000-5048.20170512
shu

焦磷酸测序技术对阿司匹林个体化用药相关SNP位点分型

  • 1.

    中国药科大学药物质量与安全预警教育部重点实验室

  • 2.

    河北省人民医院药学部

  • 3.

    南京军区南京总医院药理科

基金项目: 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

Genotype method for SNPs related to individualized aspirin treatment based on pyrosequencing technology

摘要

    摘要
  • 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 are significant individual differences in the antiplatelet effects of aspirin. Three single nucleotide polymorphisms(SNPs), rs5918, rs12041331 and rs730012, are reported to significantly correlate with the efficacy and side effects of aspirin. In the present study, the genotyping method of the three SNPs was established based on the combination of polymerase chain reaction and pyrosequencing technology. Amplification and sequencing primers were designed independently; the amplification conditions were optimized to amplify the three SNPs in the same condition. The sensitivity of the method was detected using original genome DNA at different concentrations. In order to testify the accuracy of the method, the proposed method and Sanger sequencing technology were both used to genotype the three SNPs in 20 blood samples. The results demonstrated that the genotyping method of aspirin-related SNPs was successfully established, with the detection limit being as low as 0. 4 ng genome DNA. The genotype results of 20 samples by the proposed method were exactly the same as that of Sanger sequencing. It is evident that the proposed method is sensitive and accurate.
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    • 参考文献(20)
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  • 刊出日期:  2017-10-24

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