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Clinical Chemistry 50: 2028-2036, 2004. First published September 13, 2004; 10.1373/clinchem.2004.035071
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Right arrow Molecular Diagnostics and Genetics
(Clinical Chemistry. 2004;50:2028-2036.)
© 2004 American Association for Clinical Chemistry, Inc.

Molecular Diagnostics and Genetics

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

Susan Bortolin1,a, Margot Black1, Hemanshu Modi1, Ihor Boszko1, Daniel Kobler1, Dan Fieldhouse1, Eve Lopes1, Jean-Michel Lacroix1, Rebecca Grimwood2, Philip Wells2, Richard Janeczko1 and Roman Zastawny1

1 Tm Bioscience Corporation, Toronto, Ontario, Canada.
2 Ottawa Health Research Institute, The Department of Medicine, University of Ottawa, Division of Hematology, Ottawa, Ontario, Canada.

aAddress correspondence to this author at: Tm Bioscience Corporation, 439 University Ave., Toronto, Ontario, Canada M5G 1Y8. Fax 416-593-1870; e-mail susanb{at}tmbioscience.com.

Background: We have developed a novel, microsphere-based universal array platform referred to as the Tag-ItTM 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 xMAPTM 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.




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