title: Analytical validation of the tag-it high-throughput microsphere-based universal array genotyping platform: application to the multiplex detection of a panel of thrombophilia-associated single-nucleotide polymorphisms.
abstract: BACKGROUND
  We have developed a novel, microsphere-based universal array platform referred to as the Tag-It platform. This platform is suitable for high-throughput clinical genotyping applications and was used for multiplex analysis of a panel of thrombophilia-associated single-nucleotide polymorphisms (SNPs).
  
  
  METHODS
  Genomic DNA from 132 patients was amplified by multiplex PCR using 6 primer sets, followed by multiplex allele-specific primer extension using 12 universally tagged genotyping primers. The products were then sorted on the Tag-It array and detected by use of the Luminex xMAP system. Genotypes were also determined by sequencing.
  
  
  RESULTS
  Empirical validation of the universal array showed that the highest nonspecific signal was 3.7% of the specific signal. Patient genotypes showed 100% concordance with direct DNA sequencing data for 736 SNP determinations.
  
  
  CONCLUSIONS
  The Tag-It microsphere-based universal array platform is a highly accurate, multiplexed, high-throughput SNP-detection platform.
authors.affiliation: Tm Bioscience Corporation, Toronto, Ontario, Canada. susanb@tmbioscience.com
authors.forename: Susan
authors.initials: S
authors.lastname: Bortolin
authors.forename: Margot
authors.initials: M
authors.lastname: Black
authors.forename: Hemanshu
authors.initials: H
authors.lastname: Modi
authors.forename: Ihor
authors.initials: I
authors.lastname: Boszko
authors.forename: Daniel
authors.initials: D
authors.lastname: Kobler
authors.forename: Dan
authors.initials: D
authors.lastname: Fieldhouse
authors.forename: Eve
authors.initials: E
authors.lastname: Lopes
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Lacroix
authors.forename: Rebecca
authors.initials: R
authors.lastname: Grimwood
authors.forename: Philip
authors.initials: P
authors.lastname: Wells
authors.forename: Richard
authors.initials: R
authors.lastname: Janeczko
authors.forename: Roman
authors.initials: R
authors.lastname: Zastawny
mesh_terms: D005838:Genotype; D006801:Humans; D008863:Microspheres; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide; D019851:Thrombophilia / Q000235:genetics*