Genetic mutations such as single nucleotide polymorphisms (SNP) are known as one of the most common forms which related to various genetic disorders and cancers. Among of the methods developed for efficient detection of such SNP, polymerase chain reaction (PCR) methods are widely used worldwide for its cost and viable advantages. However, the technique to discriminate small amounts of SNP mixed in abundant normal DNA is incomplete due to intrinsic technical problems of PCR such as amplification occurring even in 3’mismatched cases because of high enzyme activity of DNA polymerases. To overcome the issue, specifically designed PCR platform, STexS (SNP typing with excellent specificity) using double stranded oligonucleotides was implemented as a means to emphasize the amplification of SNP templates by decreasing unwanted amplification of 3’mismatched DNA copies. In this study, the results indicate several EGFR mutations were easily detected specifically utilizing the STexS platform. Further trials show the novel method works effectively to discriminate mutations in not only general allele specific (AS)-PCRs, but also amplification refractory mutation system (ARMS)-PCR. The STexS platform will give aid in PCRs targeting potential SNPs or genetically mutated biomarkers in human clinical samples.

单核苷酸多态性 (SNP) 等基因突变被认为是与各种遗传疾病和癌症相关的最常见形式之一。在为有效检测此类 SNP 而开发的方法中，聚合酶链反应 (PCR) 方法因其成本和可行的优势而在世界范围内广泛使用。然而，由于 DNA 聚合酶的高酶活性，即使在 3' 错配的情况下也会发生扩增等固有的 PCR 技术问题，因此区分混合在丰富的正常 DNA 中的少量 SNP 的技术是不完整的。为了克服这个问题，专门设计的PCR平台，STexS（小号NP牛逼yping与前cellent小号特异性) 使用双链寡核苷酸作为一种手段，通过减少 3' 错配 DNA 拷贝的不需要的扩增来强调 SNP 模板的扩增。在这项研究中，结果表明使用 STexS 平台可以轻松检测到几个 EGFR 突变。进一步的试验表明，这种新方法不仅可以有效区分一般等位基因特异性 (AS)-PCR 中的突变，还可以有效区分扩增难治性突变系统 (ARMS)-PCR。STexS 平台将有助于针对人类临床样本中潜在 SNP 或基因突变生物标志物的 PCR。