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Vol. 9, Issue 10, 936-949, October 1999

LETTER
A Multilocus Genotyping Assay for Candidate Markers of Cardiovascular Disease Risk

Suzanne Cheng,1,9 Michael A. Grow,1 Céline Pallaud,2 William Klitz,3 Henry A. Erlich,1 Sophia Visvikis,2 John J. Chen,3,4 Clive R. Pullinger,5 Mary J. Malloy,6,7 Gérard Siest,2 and John P. Kane6,8

1 Department of Human Genetics, Roche Molecular Systems, Inc., Alameda, California 94501 USA; 2 Centre de Médecine Préventive, 54501 Vandoeuvre-lès-Nancy, France; 3 School of Public Health and 4 Department of Integrative Biology, University of California at Berkeley, Berkeley, California 94720 USA; 5 Cardiovascular Research Institute, 6 Department of Medicine, 7 Department of Pediatrics, and 8 Department of Biochemistry and Biophysics, University of California at San Francisco, San Francisco, California 94143 USA

A number of chronic diseases, including cardiovascular disease, appear to have a multifactorial genetic risk component. Consequently, techniques are needed to facilitate evaluation of complex genetic risk factors in large cohorts. We have designed a prototype assay for genotyping a panel of 35 biallelic sites that represent variation within 15 genes from biochemical pathways implicated in the development and progression of cardiovascular disease. Each DNA sample is amplified using two multiplex polymerase chain reactions, and the alleles are genotyped simultaneously using an array of immobilized, sequence-specific oligonucleotide probes. This multilocus assay was applied to two types of cohorts. Population frequencies for the markers were estimated using 496 unrelated individuals from a family-based cohort, and the observed values were consistent with previous reports. Linkage disequilibrium between consecutive pairs of markers within the apoCIII, LPL, and ELAM genes was also estimated. A preliminary analysis of single and pairwise locus associations with severity of atherosclerosis was performed using a composite cohort of 142 individuals for whom quantitative angiography data were available; evaluation of the potentially interesting associations observed will require analysis of an independent and larger cohort. This assay format provides a research tool for studies of multilocus genetic risk factors in large cardiovascular disease cohorts, and for the subsequent development of diagnostic tests.

4   Present address: School of Public Health, St. Louis University, St. Louis, Missouri 63123 USA.
9   Corresponding author.

9:936-949 ©1999 by Cold Spring Harbor Laboratory Press  ISSN 1088-9051/99 $5.00
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