title: High-throughput genotyping of single nucleotide polymorphisms using new biplex invader technology.
abstract: The feasibility of large-scale genome-wide association studies of complex human disorders depends on the availability of accurate and efficient genotyping methods for single nucleotide polymorphisms (SNPs). We describe a new platform of the invader assay, a biplex assay, where both alleles are interrogated in a single reaction tube. The assay was evaluated on over 50 different SNPs, with over 20 SNPs genotyped in study cohorts of over 1500 individuals. We assessed the usefulness of the new platform in high-throughput genotyping and compared its accuracy to genotyping results obtained by the traditional monoplex invader assay, TaqMan genotyping and sequencing data. We present representative data for two SNPs in different genes (CD36 and protein tyrosine phosphatase 1beta) from a study cohort comprising over 1500 individuals with high or low-normal blood pressure. In this high-throughput application, the biplex invader assay is very accurate, with an error rate of <0.3% and a failure rate of 1.64%. The set-up of the assay is highly automated, facilitating the processing of large numbers of samples simultaneously. We present new analysis tools for the assignment of genotypes that further improve genotyping success. The biplex invader assay with its automated set-up and analysis offers a new efficient high-throughput genotyping platform that is suitable for association studies in large study cohorts.
authors.affiliation: Stanford Human Genome Center, Stanford University School of Medicine, 975 California Avenue, Palo Alto, CA 94305, USA. molivier@mcw.edu
authors.forename: Michael
authors.initials: M
authors.lastname: Olivier
authors.forename: Lee-Ming
authors.initials: LM
authors.lastname: Chuang
authors.forename: Mau-Song
authors.initials: MS
authors.lastname: Chang
authors.forename: Ying-Tsung
authors.initials: YT
authors.lastname: Chen
authors.forename: Dee
authors.initials: D
authors.lastname: Pei
authors.forename: Koustubh
authors.initials: K
authors.lastname: Ranade
authors.forename: Anniek
authors.initials: A
authors.lastname: de Witte
authors.forename: Jennifer
authors.initials: J
authors.lastname: Allen
authors.forename: Nguyet
authors.initials: N
authors.lastname: Tran
authors.forename: David
authors.initials: D
authors.lastname: Curb
authors.forename: Richard
authors.initials: R
authors.lastname: Pratt
authors.forename: Henk
authors.initials: H
authors.lastname: Neefs
authors.forename: Monika
authors.initials: M
authors.lastname: de Arruda Indig
authors.forename: Scott
authors.initials: S
authors.lastname: Law
authors.forename: Bruce
authors.initials: B
authors.lastname: Neri
authors.forename: Lu
authors.initials: L
authors.lastname: Wang
authors.forename: David R
authors.initials: DR
authors.lastname: Cox
mesh_terms: D001709:Biotechnology / Q000379:methods; D018955:CD36 Antigens / Q000235:genetics; D016000:Cluster Analysis; D015331:Cohort Studies; D005838:Genotype; D006801:Humans; D020641:Polymorphism, Single Nucleotide; D017027:Protein Tyrosine Phosphatases / Q000235:genetics; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D018955:CD36 Antigens; D017027:Protein Tyrosine Phosphatases