Analytical Sciences

Genotyping of the Human Lipoprotein Lipase Gene by Ferrocenylnaphthalene Diimide-based Electrochemical Hybridization Assay

Takahiko NOJIMA,*¹ Kenichi YAMASHITA,*¹ Atsuko TAKAGI,*² Yasuyuki IKEDA,*³ Hiroki KONDO,*⁴ and Shigeori TAKENAKA*⁵

*1 Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Fukuoka 819-0395, Japan
*2 Department of Etiology and Pathophysiology, National Cardiovascular Center Research Institute,Osaka 565-8565, Japan
*3 Department of Pharmacology, National Cardiovascular Center Research Institute, Osaka 565-8565, Japan
*4 Department of Biochemical Engineering and Science, Kyushu Institute of Technology, Iizuka 820-8502, Japan
*5 Department of Materials Science, Faculty of Engineering, Kyushu Institute of Technology, Kitakyushu 804-8550, Japan

A ferrocenylnaphthalene diimide (FND)-based electrochemical hybridization assay (FND-EHA) was applied to the detection of two mutations in human lipoprotein lipase (LPL) gene, G188E (one base transition) and Arita (one base deletion). A probe oligodeoxyribonucleotide of 13 bases representing the wild type (WT) sequence of LPL was immobilized on a gold electrode, followed by hybridization with a sample PCR product of 350 base pairs under conditions in which both WT and mutated (MT) sequences could form a duplex with the probe. The hybridized electrodes were soaked in an electrolyte containing FND under conditions in which only the mismatched duplex could undergo dissociation. FND was concentrated in proportion to the amount of the duplex remaining on the electrode to give rise to a current signal. Blind tests were run to judge the genotype (WT/WT, WT/MT, or MT/MT) of 10 samples each for the G188E and Arita mutations and then, 8 and 10 of them were judged correctly, respectively.

J-STAGE: View this article in J-STAGE