Multiplex SNP analysis on whole genome amplified DNA from archived dried bloodspots, a validation study Kristine C. Tvedegaard,1 Erik Parner,1 Craig W. Hooper,2 Jørn Atterman,1 Niels Gregersen3, Poul Thorsen,1 1Institute of Public Health, NANEA at Department of Epidemiology, University of Aarhus, Aarhus, Denmark; 2Hereditary Blood Disorders, National Centers for Birth Defects and Developmental Disabilities, Centers for Disease Control and Prevention, Atlanta GA, United States; 3Reseach Unit for Molecular Medicine, Skejby Sygehus, Aarhus, Denmark BACKGROUND: The EraGen Multi-Code System is a further development of allele specific primer extension (ASPE) for multiplex SNP analysis based on the Luminex 100 IS platform. It uses isobases (isoC and isoG) and the software MultiCode-PLx platform for data analysis and data handling. We validate the EraGen multicode system in two 6-plex assays used on whole genome amplified (WGA) DNA from archived dried bloodspots. METHODS AND MATERIAL: The chemically synthesized new base pair (isoC and isoG) allow for the creation of new DNA strands that provide superior specificity and allow development of assays with greater sensitivity than conventional methods. To validate the method 900 WGA DNA samples were genotyped in duplets. 10-20 % of all samples were sequenced to be used as reference. Accuracy and repeatability was estimated for each SNP. Robustness was estimated as rate of conclusive outcomes for each SNP. RESULTS: The accuracy ranged from 98.3-100%, repeatability ranged from 99.2-99.7% and robustness ranged from 94.1-99.3%. CONCLUSION: The Multi-Code System is a highly sensitive and specific method for multiplex SNP analysis on WGA DNA from archived dried bloodspots.
Conference: Planet Xmap, Los Angeles, Usa, 12 - 14 March 2007, 2007