本文選題：GLPR基因 + 高分辨率熔解曲線　； 參考：《中國(guó)藥房》2017年17期

【摘要】：目的:建立檢測(cè)腸促胰島素樣肽-1受體(GLP1R)基因已知突變位點(diǎn)rs3765467(NT_007592.16的第39 065 819位)的方法,并評(píng)價(jià)其準(zhǔn)確性與實(shí)用性。方法:收集我院體檢中心2015年10月-2016年2月72例健康體檢者的外周靜脈血樣本,采用柱提法提取其全血DNA,經(jīng)降落聚合酶鏈反應(yīng)擴(kuò)增后,采用高分辨率熔解曲線(HRM)法對(duì)產(chǎn)物進(jìn)行分析;同時(shí)選取其中38例受試樣本進(jìn)行雙脫氧鏈終止法(Sanger測(cè)序法)測(cè)序驗(yàn)證,比較2種方法的結(jié)果。結(jié)果:突變掃描結(jié)果顯示,擴(kuò)增片段中存在39 065 817和39 065 819兩個(gè)多態(tài)性位點(diǎn)。HRM法只檢測(cè)出了4種基因型[GCG/GCG、GCA/GCG或ACG/GCG、GCA/GCA或ACG/ACG、A(G)CA(G)];而Sanger測(cè)序法共檢測(cè)出6種基因型[GCG/GCG、ACG/GCG、ACG/ACG、A(G)CA(G)、GCA/GCG、GCA/GCA]。結(jié)論:HRM法可區(qū)分GCG/GCG和A(G)CA(G)基因型,但無(wú)法區(qū)分GCA/GCG與ACG/GCG雜合突變、GCA/GCA與ACG/ACG純合突變。該方法并不適用于多個(gè)鄰近位點(diǎn)的單核苷酸多態(tài)性檢測(cè)。在進(jìn)行單核苷酸突變檢測(cè)時(shí),應(yīng)對(duì)序列進(jìn)行綜合分析后再選取經(jīng)濟(jì)、簡(jiǎn)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.
【作者單位】： 四川省醫(yī)學(xué)科學(xué)院/四川省人民醫(yī)院藥學(xué)部;
【基金】：四川省科技支撐計(jì)劃項(xiàng)目(No.2015SZ0182)
【分類號(hào)】：R969
，

本文編號(hào)：1906252