Validation of the Loop-Mediated Isothermal Amplification Method for Single Nucleotide Polymorphism Genotyping with Whole Blood

Authors: Iwasaki, Masaomi¹; Yonekawa, Toshihiro²; Otsuka, Kimihiko²; Suzuki, Wataru²; Nagamine, Kentaro²; Hase, Tetsu²; Tatsumi, Ke-Ita³; Horigome, Tsuneyoshi⁴; Notomi, Tsugunori²; Kanda, Hidetoshi²

Source: Genome Letters, Volume 2, Number 3, September 2003 , pp. 119-126(8)

Publisher: American Scientific Publishers

Buy & download fulltext article:

Price: $113.00 plus tax (Refund Policy)

Abstract:

We applied the loop-mediated isothermal amplification (LAMP) method to single nucleotide polymorphism (SNP) typing of the drug-responsive cytochrome P450 CYP2C19 gene. The CYP2C19*1, CYP2C19*2, and CYP2C19*3 alleles of the cytochrome P450 gene were easily and completely distinguished from each other even in the presence of homologous genes by means of allele-specific LAMP. The products of the LAMP reaction, which was performed in the presence of an intercalating dye, were detected within 30 min without any post-reaction sample manipulation. When whole blood was used as the specimen, LAMP-based SNP typing required only heat treatment, and purification of genomic DNA from the specimen was unnecessary. In addition, the products of the LAMP reaction were detected within 30 min, as was the case when genomic DNA was used as the sample. In a study of human blood samples, the results obtained by the LAMP method completely agreed with those obtained by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. LAMP-based SNP typing is an accurate, rapid, and simple method that may be useful for SNP typing, especially for point-of-care testing.

Keywords: LAMP; SNP; GENOTYPING; POCT; DNA AMPLIFICATION; DRUG-RESPONSIVE GENE; P450

Document Type: Research article

DOI: http://dx.doi.org/10.1166/gl.2003.028

Affiliations: 1: Eiken Chemical Co., Ltd., Tochigi, Japan and Courses of Biosphere Science and Functional Biology, Graduate School of Science and Technology, Niigata University, Japan 2: Eiken Chemical Co., Ltd., Tochigi, Japan 3: Osaka University, Graduate School of Medicine, Osaka, Japan 4: Courses of Biosphere Science and Functional Biology, Graduate School of Science and Technology, Niigata University, Japan

Publication date: 2003-09-01

More about this publication?

Editorial Board
Information for Authors
ingentaconnect is not responsible for the content or availability of external websites

Validation of the Loop-Mediated Isothermal Amplification Method for Single Nucleotide Polymorphism Genotyping with Whole Blood

Buy & download fulltext article:

Tools

Key

Text size:

Useful pages