Application of Denaturing HPLC for Detection of G/C Polymorphism in the Cyclo-Oxygenase-2 Gene

作者：Huuskonen K H; Kunnas T A; Tanner M M; Kortelainen M L; Nikkari S T^*

来源：Journal of Liquid Chromatography & Related Technologies, 2009, 32(2): 201-209.

DOI：10.1080/10826070802602973

• Cyclo-oxygenase-2 gene
• Denaturing HPLC
• DHPLC
• G/C Polymorphism

Denaturing high performance liquid chromatography (DHPLC) is a fast and sensitive method to detect gene mutations. The aim of this study was to examine whether DHPLC is an appropriate method to detect a G/C transversion in the promoter region of human cyclo-oxygenase-2 gene. A fragment of 228 bp was amplified from 336 subjects for DHPLC analysis. Results were confirmed by restriction endonuclease analysis. Considering that detection of G/C transversion is quite challenging by DHPLC, the specificity was high ( 94,3%). Nonetheless, resolution significantly decreased after 400 samples, while it should be possible to analyze 1000 samples per column.
According to our findings, DHPLC is not the most adequate method to detect G/C transversion in the gene because of the relatively rapid loss of column resolution. This should be kept in mind when genomic DNA is screened for unknown mutations.

• 出版日期2009
• 单位河北医科大学