采用改进的三引物等位基因扩增法对痛风相关SNP位点的分型研究  

Research on gout-related SNP genotyping by improved tri-primer allele gene amplification

作　　者：郑晨娜 王清瑶[2] 黄玉香 叶桂华[3] 许超尘 杨会勇[2] 

机构地区：[1]福建省泉州市医学高等专科学校,福建泉州362000 [2]福建省泉州市华侨大学分子药物研究院,福建泉州362000 [3]福建省泉州市华侨大学校医院检验科,福建泉州362000 [4]福建省泉州正骨医院,福建泉州362000

出　　处：《重庆医学》2014年第12期1476-1479,共4页Chongqing medicine

基　　金：福建省教育厅A类资助项目(JA11323);泉州市科技局重点资助项目(2011Z5;2011Z30)

摘　　要：目的对三引物等位基因扩增法进行改进,实现直接对外周血样本进行痛风相关单核苷酸多态性(SNP)位点分型。方法针对乙二胺四乙酸(EDTA)、肝素钠和枸橼酸钠等临床常用抗凝处理的外周血样品,以rs1165205位点为靶位点,配制适用于全血扩增的"YW"缓冲液,优化聚合酶链反应(PCR)扩增体系和扩增条件,实现SNP分型检测。选取40例男性健康志愿者和40例痛风患者,对其进行SNP基因型检测。结果改进后的三引物等位基因扩增法对rs1165205位点分型结果与Sanger测序检测一致。80个样本中rs1165205位点各基因型在发病人群和未发病人群分布差异无统计学意义(P=0.335)。结论改进后的三引物等位基因特异性扩增法可以直接对临床常用抗凝处理的外周血样品痛风SNP位点进行快速分型研究。Objective To improve the tri-primer allele gene amplification for realizing the single nucleotide polymorphisms （SNP） genotyping of the peripheral blood sample .Methods Aiming at the peripheral blood samples with the clinical usual antico-agulation processing by EDTA ,heparin and citrate ,with the locus rs1165205 as the target site ,the buffer solution（YW） suitable for whole blood was prepared and the PCR amplification system and the amplification condition were optimized for realizing the detec-tion of SNP genotyping .Results The genotyping results of locus rs1165205 by improved tri-primer allele gene amplification method were consistent with the results of the Sanger sequencing method ,and the peripheral blood samples treated by different anticoagu-lant were genotyped by the improved tri-primer ASA .Among 80 samples ,various genotypes of locus rs1165205 had no statistical differences in the distribution between the gout population and non-gout population（P= 0 .335） .Conclusion The improved tri-primer allele gene amplification method can be adopted to conduct the rapid genotyping research on gout SNP locus of the peripheral blood samples with the clinical usual anticoagulation processing .

关 键 词：三引物扩增法 等位基因 基因扩增 痛风 多态性  单核苷酸 

分 类 号：R450[医药卫生—治疗学]