高分辨率熔解曲线法在GLP1R基因多态性位点rs3765467检测中的应用

发布时间：2018-05-18 14:51

  本文选题：GLPR基因 + 高分辨率熔解曲线　； 参考：《中国药房》2017年17期

【摘要】：目的:建立检测肠促胰岛素样肽-1受体(GLP1R)基因已知突变位点rs3765467(NT_007592.16的第39 065 819位)的方法,并评价其准确性与实用性。方法:收集我院体检中心2015年10月-2016年2月72例健康体检者的外周静脉血样本,采用柱提法提取其全血DNA,经降落聚合酶链反应扩增后,采用高分辨率熔解曲线(HRM)法对产物进行分析;同时选取其中38例受试样本进行双脱氧链终止法(Sanger测序法)测序验证,比较2种方法的结果。结果:突变扫描结果显示,扩增片段中存在39 065 817和39 065 819两个多态性位点。HRM法只检测出了4种基因型[GCG/GCG、GCA/GCG或ACG/GCG、GCA/GCA或ACG/ACG、A(G)CA(G)];而Sanger测序法共检测出6种基因型[GCG/GCG、ACG/GCG、ACG/ACG、A(G)CA(G)、GCA/GCG、GCA/GCA]。结论:HRM法可区分GCG/GCG和A(G)CA(G)基因型,但无法区分GCA/GCG与ACG/GCG杂合突变、GCA/GCA与ACG/ACG纯合突变。该方法并不适用于多个邻近位点的单核苷酸多态性检测。在进行单核苷酸突变检测时,应对序列进行综合分析后再选取经济、简便的方法。
[Abstract]:Aim: to establish a method for the detection of the 39,065,819 known mutation site of the intestinal insulin-stimulating peptide 1 receptor (GLP1R) gene, and to evaluate its accuracy and practicability. Methods: the peripheral venous blood samples were collected from 72 healthy persons from October 2015 to February 2016 in our hospital. The whole blood DNA was extracted by column extraction and amplified by landing polymerase chain reaction (PCR). The products were analyzed by high resolution melting curve (HRM) method, and 38 samples were tested by dideoxy chain termination method (Sanger sequencing) and the results of the two methods were compared. Results: the results of mutation scanning showed that there were two polymorphic loci (39 065 817 and 39 065 819) in the amplified fragments. Only four genotypes were detected by HRM. Conclusion the GCG/GCG gene can be distinguished from the Agna GCAG genotype by the method of: HRM, but the heterozygous mutation of GCA/GCG and ACG/GCG can not be distinguished from the homozygous mutation of GCA / GCA and ACG/ACG. This method is not suitable for the detection of single nucleotide polymorphisms at multiple adjacent loci. In the detection of single nucleotide mutation, the economic and simple method should be selected after comprehensive analysis of the sequence.
【作者单位】： 四川省医学科学院/四川省人民医院药学部;
【基金】：四川省科技支撑计划项目(No.2015SZ0182)
【分类号】：R969
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本文编号：1906252