Sanger测序和AMRS-PCR检测ALDH2基因突变结果的比较

Comparison of Sanger sequencing and ARMS-PCR in the detection of ALDH2 gene mutation

目的比较Sanger测序法与扩增阻滞突变系统PCR(ARMS-PCR)在检测患者线粒体乙醛脱氢酶2(ALDH2)基因突变方面的差异。方法将2019年10月至2020年1月于陆军军医大学第一附属医院治疗的350例有冠心病、心肌梗死、心绞痛、原发性高血压病史的患者纳入研究,收集受试者外周血标本。用Sanger测序法与ARMS-PCR法检测ALDH2基因突变,分析两种方法检测结果的差异。结果两种方法检测ALDH2基因突变的总突变率均为33.1%,AA纯合型突变率为3.7%,GA杂合型突变率为29.4%。两种方法检测结果的总符合率为100.00%,突变型阳性符合率和阴性符合率均为100.00%,ARMS-PCR法与Sanger测序法对临床标本的检测结果一致性较好(Kappa≥0.7,P<0.05),具有等效性。另外,ARMS-PCR法检测出的ALDH2基因型分布情况与性别、年龄无关(P>0.05)。结论Sanger测序法和ARMS-PCR法均可准确、有效地检测ALDH2突变情况,但基于成本优势和操作简便的原则,ARMS-PCR法在临床实践中更具优势。

Objective To compare the differences between Sanger sequencing method and amplification refractory mutation system PCR(ARMS-PCR)in detecting mitochondrial aldehyde dehydrogenase 2(ALDH2)gene mutations in patients.Methods A total of 350 patients with a history of coronary heart disease,myocardial infarction,angina pectoris,and essential hypertension who were treated in the First Affiliated Hospital of Army Medical University from October 2019 to January 2020 were enrolled in the study,and peripheral blood samples were collected from the subjects.ALDH2 mutations were detected by Sanger sequencing and ARMS-PCR,and the results were compared between the two methods.Results The total mutation rates of ALDH2 gene detected by two methods were both 33.1%,AA homozygous mutation rate was 3.7%,GA heterozygous mutation rate was 29.4%.The total coincidence rate of the results of the two methods was 100.00%,and the positive coincidence rate and negative coincidence rate of mutation were both 100.00%.The results of ARMS-PCR and Sanger sequencing in clinical samples were consistent(Kappa≥0.7,P<0.05),which was equivalent.In addition,the distribution of ALDH2 genotypes detected by ARMS-PCR was not related to gender and age(P>0.05).Conclusion Both Sanger sequencing and ARMS-PCR can detect ALDH2 mutations accurately and effectively,but ARMS-PCR is more advantageous in clinical practice based on cost advantage and ease of operation.