Mitani Yasumasa, Lezhava Alexander, Kawai Yuki, Kikuchi Takeshi, Oguchi-Katayama Atsuko, Kogo Yasushi, Itoh Masayoshi, Miyagi Toru, Takakura Hideki, Hoshi Kanako, Kato Chiaki, Arakawa Takahiro, Shibata Kazuhiro, Fukui Kenji, Masui Ryoji, Kuramitsu Seiki, Kiyotani Kazuma, Chalk Alistair, Tsunekawa Katsuhiko, Murakami Masami, Kamataki Tetsuya, Oka Takanori, Shimada Hiroshi, Cizdziel Paul E, Hayashizaki Yoshihide
Genome Exploration Research Group (Genome Network Project Core Group), RIKEN Genomic Sciences Center (GSC), RIKEN Yokohama Institute, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan.
Nat Methods. 2007 Mar;4(3):257-62. doi: 10.1038/nmeth1007. Epub 2007 Feb 18.
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.
我们开发了一种名为智能扩增过程版本2（SMAP 2）的快速单核苷酸多态性（SNP）检测系统。由于DNA扩增仅在引物完全匹配时发生，因此仅扩增就足以识别目标等位基因。为了实现支持这一说法所需的保真度，我们使用了两种新的互补方法来抑制由错配引发事件导致的指数背景DNA扩增。SMAP 2是等温的，当使用从嗜酸嗜热栖热菌克隆和分离的新型DNA聚合酶（Aac pol）进行操作时，可在30分钟内从全血中实现SNP检测。此外，为了帮助科学界配置SMAP 2检测方法，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，一种高精度、快速的DNA扩增技术可用于辅助药物基因组学研究和分子诊断应用。
