A New Technique—Simultaneously Detecting Multiple Genetic Polymorphisms of Type 2 Diabetes- associated Genes by the Enzymatic Chip Array

Abstract

Type 2 diabetes mellitus (T2DM) is a polygenetic disease. Its incidence is increasing continuously in Taiwan as the standard of living improves. Current diabetes research is striving to identify those high at risk of T2DM through T2DM- associated gene studies. A number of techniques are available for the molecular detection of T2DM-associated genes, including polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), direct sequencing, and TaqMan genotyping. However, they are capable of analyzing only one molecular target in each experiment. In the present study, we selected eight candidate genes that are potentially associated with T2DM [angiotensinogen (AGT), sulfonylurea receptor-1 (SUR-1), peroxisome proliferators-activated receptor-γ (PPAR-γ), PPAR-γ coactivator-1 (PGC-1), calpain-10 (CAPN10), β2-adrenergic receptor (ADRB2), mannose-binding lectin (MBL2), and insulin receptor substrate-1 (IRS-1)]. We used enzymatic chip array technology, which we had previously established, and analyzed its relevance to diabetes research. We enrolled 1280 Taiwanese patients (700 with T2DM and 580 non-diabetic controls). The genes of all subjects were analyzed by the enzymatic chip array. The results were consistent with direct sequencing. On the basis of multivariate logistic-regression analysis—with adjustment for age—the following variables were associated with a significant risk of T2DM: body mass index, serum cholesterol, triglyceride level, low- and high-density lipoprotein cholesterol levels, and polymorphisms, including PGC-1 Gly482Ser, SUR1 Arg1273Arg, ADRB2 Arg16Gly, CAPN10 SNP43, AGT Met235Thr, and MLB2 Gly54Asp. The enzymatic chip array is a useful tool for multiple gene analysis in diabetes.