Rapid SNP diagnostics using asymmetric isothermal amplification and a new mismatch-suppression technology
Rapid SNP diagnostics using asymmetric isothermal amplification and a new mismatch-suppression technology
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DOI：
10.1038/nmeth1007
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发表时间：
2007-03-01
期刊：
影响因子：
48
通讯作者：
Hayashizaki, Yoshihide
中科院分区：
文献类型：
--
作者：
Mitani, Yasumasa;Lezhava, Alexander;Hayashizaki, Yoshihide
我们开发了一种快速单核苷酸多态性 (SNP) 检测系统，名为智能扩增过程版本 2 (SMAP 2)。由于 DNA 扩增仅在引物完美匹配的情况下发生，因此仅扩增就足以识别目标等位基因。为了达到支持这一主张所需的保真度，我们使用了两种新的互补方法来抑制由错误引发事件导致的指数背景 DNA 扩增。 SMAP 2 是等温的，使用从酸热脂环酸芽孢杆菌 (Aac pol) 中克隆和分离的新型 DNA 聚合酶时，可在 30 分钟内对人全血进行 SNP 检测。此外，为了协助科学界配置 SMAP 2 检测，我们开发了专门用于 SMAP 2 引物设计的软件。有了这些新工具，高精度、快速的 DNA 扩增技术就可以帮助药物基因组学研究和分子诊断应用。
We developed a rapid single nucleotide polymorphism (SNP) detection system named smart amplification process version 2 (SMAP 2). Because DNA amplification only occurred with a perfect primer match, amplification alone was sufficient to identify the target allele. To achieve the requisite fidelity to support this claim, we used two new and complementary approaches to suppress exponential background DNA amplification that resulted from mispriming events. SMAP 2 is isothermal and achieved SNP detection from whole human blood in 30 min when performed with a new DNA polymerase that was cloned and isolated from Alicyclobacillus acidocaldarius (Aac pol). Furthermore, to assist the scientific community in configuring SMAP 2 assays, we developed software specific for SMAP 2 primer design. With these new tools, a high-precision and rapid DNA amplification technology becomes available to aid in pharmacogenomic research and molecular-diagnostics applications.