Full Text (PDF)
The RIKEN Yokohama Institute is a world renowned research center. With a focus on life science, a goal of this government institute is to improve medical, environmental, and food conditions that plague humankind. As one of the four main groups within the institute, the Genome Science Center (GSC) is conducting comprehensive structural and functional analysis of genes and proteins from their molecular nature to variations at the individual level. Within the GSC, our lab is focused on development of rapid genotyping systems to detect DNA variations with extreme precision. Improvements to the smart amplification process (SMAP) are continuously being pursued. Furthermore, through collaborations with industry and clinical partners, the technology is being applied for personalized medicine, lifestyle genetic counseling, point of care testing, infectious disease diagnostics, and food and environmental testing.
www.gsc.riken.jp/eng/group/genome/index.html
The Technique
SMAP is an isothermal technique employing a unique mismatch suppression technology and primer design that effectively eliminates background, giving extreme precision in the amplification process. The technology is capable of amplification and detection directly from lysed human blood samples without DNA purification, uses <1 µL per assay, and takes only 30–60 min from start to finish. Seminal research data has shown that SMAP has sensitivity equivalent to PCR; there are also several examples of accurately detecting single nucleotide polymorphisms (SNPs) and discriminating them from the wild-type sequence and related gene family members. A web site has been established, www.smapdna.com, with links to primer design software and other technical support for the practice of SMAP technology in basic research applications. We reported the use of a competitive probe (CP) in the SMAP-2 assay and demonstrated the complete suppression of background amplification, which enabled the accurate and rapid genotyping of the UGT1A1*28 microsatellite polymorphism. This is the first report of a CP effectively applied to an isothermal amplification technique.
See “Use of a competitive probe in assay design for genotyping of the UGT1A1*28 microsatellite polymorphism by the smart amplification process” on page 479.
