Detection of C677T mutation in methylenetetrahydrofolate reductase gene by denaturing high performance liquid chromatography

作者：Fang, NH^*; Lin, L; Ren, JC; Wu, D

来源：Biomedical Chromatography, 2004, 18(9): 625-629.

DOI：10.1002/bmc.364

• methylenetetrahydrofolate reductase (MTHFR)
• mutation detection
• denaturing high-performance chromatography
• (DHPLC)

In this paper, we described an assay for the detection of the C677T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene using denaturing high-performance liquid chromatography (DHPLC). The conditions for DHPLC analysis were systematically investigated based on a general HPLC instrument (Prostar VARIAN). A 225 by DNA fragment covering the 677 site of MTHFR gene was amplified by PCR technology using the purified DNA from whole blood or whole blood as template DNA. PCR products were directly injected without the need for purification. The C677T mutation could be clearly distinguished by DHPLC technology. Our data demonstrated that DHPLC was a powerful and alternative tool for detection of genetic variants and single-nucleotide polymorphisms to electrophoresis technology.

• 出版日期2004-11
• 单位上海交通大学