Detection of Six Single-Nucleotide Polymorphisms Associated with Rheumatoid Arthritis by a Loop-Mediated Isothermal Amplification Method and an Electrochemical DNA Chip

详细信息    查看全文

• 作者：Naoko Nakamura ; Keiko Ito ; Masayoshi Takahashi ; Koji Hashimoto ; Manabu Kawamoto ; Mariko Yamanaka ; Atsuo Taniguchi ; Naoyuki Kamatani ; Nobuhiro Gemma
• 刊名：Analytical Chemistry
• 出版年：2007
• 出版时间：December 15, 2007
• 年：2007
• 卷：79
• 期：24
• 页码：9484 - 9493
• 全文大小：385K
• 年卷期：v.79,no.24(December 15, 2007)
• ISSN：1520-6882

文摘

An electrochemical DNA chip using an electrochemicallyactive intercalator and DNA probe immobilized on a goldelectrode has been developed for genetic analysis. In thisstudy, the six polymorphisms associated with rheumatoidarthritis (RA), N-acetyltransferase2 (NAT2) gene polymorphisms T341C, G590A, and G857A, methylenetetrahydrofolate reductase (MTHFR) gene polymorphismsC677T and A1298C, and serum amyloid A1 (SAA1) genepromoter polymorphism C-13T were simultaneously detected by the electrochemical DNA chip and the loop-mediated isothermal amplification (LAMP) method, whichis a novel technique for DNA amplification. Humangenomic DNAs were extracted from blood, and the targetscontaining the six polymorphisms were amplified by theLAMP method. A sample containing the six LAMP products was reacted with the electrochemical DNA chip usinga DNA detection system that controls hybridization reaction, washing, electrochemical detection, and data analysis automatically. A total of 31 samples were genotypedby this method, and the results were completely consistentwith those determined by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)analysis or the PCR direct sequence analysis. The timerequired for this method was only 2 h, and operationswere very simple. Therefore, this method is expected tocontribute to personalized medicine based on genotype.