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Abstract
European Congress of Clinical Chemistry and Laboratory Medicine(2001.5.26〜31 ; Prague)
Loop-mediated isothermal amplification (LAMP) of DNA
Notomi Tsugunori
(Eiken Chemical Co., Ltd.)
Nucleic acid amplification is one of the most valuable tools in virtually all life science fields including application-oriented fields, such as clinical medicine, in which diagnosis of infectious disease, genetic disorder and genetic traits is particularly benefited by this new technique.
We have developed a novel method termed "Loop-mediated isothermal amplification (LAMP)". LAMP relies on auto-cycling strand displacement DNA synthesis primed by specially designed primers and amplifies DNA 3-fold per half cycle. Consequently, LAMP is highly selective to target DNA and highly efficient in amplification, enabling detection of a few copies of target DNA in less than an hour.
LAMP method has many features as follows: 1) LAMP amplifies DNA under an isothermal condition. 2) LAMP is highly specific for the target sequence due to its recognition of six independent sequences in the target DNA. 3) LAMP amplifies DNA with such a high efficiency as enabling detection of only a few copies of DNA in a short time. 4) LAMP also amplifies RNA target simply by adding reverse transcriptase. 5) Because LAMP products reach more than 0.5 mg/ml, the DNA synthesis reaction byproduct pyrophosphate that binds with magnesium, produces turbidity. This turbidity can be detected by naked eye. Also, when a fluorescent intercalater is added in the reaction mixture before reaction, the amplification products can be easily observed under an UV lamp. 6) Because LAMP discriminates a single nucleotide change at each cycling step of the DNA replication, it is very useful method for detection of SNPs of human genome.
Thus, this method is a superior alternative to other amplification methods and therefore would be useful in gene diagnostics.

European Congress of Clinical Chemistry and Laboratory Medicine(2001.5.29; Prague)

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