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Clinical Chemistry 0: clinchem.2004.035071v1, 2004; 10.1373/clinchem.2004.035071
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Received on March 31, 2004
Accepted on August 27, 2004

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 Bortolin 1*, Margot Black 1, Hemanshu Modi 1, Ihor Boszko 1, Daniel Kobler 1, Dan Fieldhouse 1, Eve Lopes 1, Jean-Michel Lacroix 1, Rebecca Grimwood 2, Philip Wells 2, Richard Janeczko 1, Roman Zastawny 1

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.

* To whom correspondence should be addressed. 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 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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