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Abstract
Cambridge Healthtech Institute's Conference
A novel SNP typing technology based on the nucleic acid amplification method, LAMP.
Hidetoshi Kanda, Masaomi Iwasaki, Toshihiro Yonekawa, Kentaro Nagamine, Yasuyoshi Mori, Norihiro Tomita, Tetsu Hase and Tsugunori Notomi
(DUG Unit, Eiken Chemical Co., Ltd.)
A variety of SNP typing technologies have so far been reported. However, more convenient and accurate methods are now required. Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method that amplifies DNA specifically and rapidly under isothermal conditions (1).
Recently we have found that specially designed primers of LAMP discriminated a single nucleotide change at each cycling step of the DNA replication. Thus, the type of SNP can easily be detected by just amplifying the DNA containing the SNP. Actually, CYP2C19*1, CYP2C19*2, CYP2C19*3 alleles of drug responsive P450 gene were easily and strictly discriminated from each other even in coexistence with their homologous genes, CYP2C9, CYP2C8, CYP2C18 and so on, by using LAMP. A blind study carried out on human genomic DNA samples showed that the method was as accurate as PCR-RFLP. The products of LAMP reactions, carried out in the presence of intercalating dye, were detected within 30 min without the need for any post-reaction sample manipulation.
LAMP-based SNP typing is an accurate, rapid, easy and inexpensive method, and accordingly the method appears useful for SNP typing at POC testing.

1)Tsugunori Notomi, Hiroto Okayama, Harumi Masubuchi, Toshihiro Yonekawa, Keiko Watanabe, Nobuyuki Amino and Tetsu Hase
" Loop-mediated isothermal amplification of DNA."
Nucleic Acids Research, 2000, Vol.28 No.12, e63

Cambridge Healthtech Institute's Conference(2001.3.26〜29;Tokyo)

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