Department of Pathology, Mackay Memorial Hospital, Taipei, Taiwan.
Department of Nuclear Medicine, Chang Gung Memorial Hospital, Taoyuan, Taiwan.
BMC Biotechnol. 2019 Aug 23;19(1):62. doi: 10.1186/s12896-019-0555-1.
In personalized medicine, companion diagnostic tests provide additional information to help select a treatment option likely to be optimal for a patient. Although such tests include several techniques for detecting low levels of mutant genes in wild-type backgrounds with fairly high sensitivity, most tests are not specific, and may exhibit high false positive rates. In this study, we describe a new primer structure, named 'stuntmer', to selectively suppress amplification of wild-type templates, and promote amplification of mutant templates.
A single stuntmer for a defined region of DNA can detect several kinds of mutations, including point mutations, deletions, and insertions. Stuntmer PCRs are also highly sensitive, being able to amplify mutant sequences that may make up as little as 0.1% of the DNA sample.
In conclusion, our technique, stuntmer PCR, can provide a simple, low-cost, highly sensitive, highly accurate, and highly specific platform for developing companion diagnostic tests.
在个性化医疗中，伴随诊断测试提供了额外的信息，以帮助选择最适合患者的治疗方案。虽然这些测试包括几种用于检测野生型背景中低水平突变基因的技术，具有相当高的灵敏度，但大多数测试并不特异，可能表现出较高的假阳性率。在这项研究中，我们描述了一种新的引物结构，命名为“stuntmer”，用于选择性地抑制野生型模板的扩增，并促进突变型模板的扩增。
针对 DNA 的特定区域的单个 stuntmer 可以检测到多种突变，包括点突变、缺失和插入。Stuntmer PCR 也非常敏感，能够扩增可能只占 DNA 样本 0.1%的突变序列。
总之，我们的技术，stuntmer PCR，可以为开发伴随诊断测试提供一个简单、低成本、高敏感、高准确、高特异的平台。
