Single nucleotide polymorphism (SNP) analysis based on allele-specific polymerase chain reaction (AS-PCR) is a relatively effective and economical method compared with other genotyping technologies such as DNA sequencing, DNA hybridization and isothermal amplification strategies. But AS-PCR is limited by its labor-intensive optimization of reaction parameters and time-consuming result assessment. In this study, we put forward a novel idea of data processing to address this problem. SNP analysis was accomplished by AS-PCR with endpoint electrochemical detection. For each sample, two separate reactions were run simultaneously with two sets of allele-specific primers (wild-type primers for W system and mutant primers for M system). We measured their redox current signals on screen-printed electrodes once AS-PCR finished and calculated the difference value of current signals between two systems to determine the genotyping result. Based on the difference value of fluorescent signals, real-time fluorescent PCR was used to study reaction parameters in AS-PCR. With screened parameters, we obtained the genotyping results within 50 min. 36 hair-root samples from volunteers were analyzed by our method and their genotypes of ALDH2 gene (encoding aldehyde dehydrogenase 2) were totally identical with data from commercialized sequencing. Our work first employed difference value between two reaction systems to differentiate allele and provided a novel idea of data processing in AS-PCR method. It is able to promote the quick analysis of SNP in the fields of health monitor, disease precaution, and personalized diagnosis and treatment.

与其他基因分型技术（例如DNA测序，DNA杂交和等温扩增策略）相比，基于等位基因特异性聚合酶链反应（AS-PCR）的单核苷酸多态性（SNP）分析是一种相对有效且经济的方法。但是AS-PCR受其劳动密集型反应参数优化和费时的结果评估的限制。在这项研究中，我们提出了一种新的数据处理方法来解决这个问题。SNP分析通过具有终点电化学检测的AS-PCR完成。对于每个样品，使用两组等位基因特异性引物（W系统的野生型引物和M系统的突变型引物）同时进行两个单独的反应。AS-PCR完成后，我们在丝网印刷电极上测量了它们的氧化还原电流信号，并计算了两个系统之间电流信号的差值，以确定基因分型结果。根据荧光信号的差值，采用实时荧光PCR研究AS-PCR中的反应参数。使用筛选的参数，我们在50分钟内获得了基因分型结果。通过我们的方法分析了36名来自志愿者的发根样品，并分析了它们的基因型。ALDH2基因（编码醛脱氢酶2）与商业测序数据完全相同。我们的工作首先利用两个反应系统之间的差异值来区分等位基因，并为AS-PCR方法中的数据处理提供了新思路。它可以促进在健康监测，疾病预防以及个性化诊断和治疗领域中SNP的快速分析。