A multilocus genotyping assay for candidate markers of cardiovascular disease risk

A number of chronic diseases, including cardiovascular disease, appear to h ave a multifactorial generic risk component. Consequently, techniques are n eeded to facilitate evaluation of complex genetic risk factors in large coh orts. We have designed a prototype assay For genotyping a panel of 35 biall elic sites that represent variation within 15 genes from biochemical pathwa ys implicated in the development and progression of cardiovascular disease. Each DNA sample is amplified using two multiplex polymerase chain reaction s, and the alleles are genotyped simultaneously using an array of immobiliz ed, sequence-specific oligonucleotide probes. This multilocus assay was app lied to two types of cohorts. Population frequencies For the markers were e stimated using 496 unrelated individuals from a Family-based cohort, and th e observed values were consistent with precious reports. Linkage disequilib rium between consecutive pairs of markers within the apoCIII, LPL, and ELAM genes was also estimated. A preliminary analysis of single and pairwise lo cus associations with severity of atherosclerosis was performed using a com posite cohort of 142 individuals For whom quantitative angiography data wer e available; evaluation of the potentially interesting associations observe d will require analysis of an independent and larger cohort. This assay for mat provides a research tool for studies of multilocus genetic risk factors in large cardiovascular disease cohorts, and for the subsequent developmen t of diagnostic tests.