[Abstract]  [Full Text]  [PDF]

Seok Hwee Koo¹, Tan Ching Ong², Kok Ting Chong¹, Caroline Guat Lay Lee³, Fook Tim Chew² and Edmund Jon Deoon Lee^1*

¹ Department of Pharmacology, National University of Singapore. Singapore.
² Department of Biological Sciences, National University of Singapore. Singapore.
³ Department of Biochemistry, National University of Singapore. Block MD11, Level 5, #05-09, Clinical Research Centre, 10 Medical Drive. Singapore 117597. Phone: +65-65168437 Fax: +65-68737690 Email: edlee@nus.edu.sg.

* To whom correspondence should be addressed: Edmund Jon Deoon Lee, Department of Pharmacology, National University of Singapore. Singapore.

Biol. Proced. Online 2007;9:27-42. doi:10.1251/bpo131
Submitted: March 12, 2007; Accepted: July 3, 2007; Published: November 1, 2007.

Indexing terms: Genotype; Microspheres; Polymorphism, Genetic.

Abbreviations: ABC, ATP-binding cassette; ASPE, allele-specific primer extension; EDC, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride; MES, 2[N-Morpholino] ethanesulfonic acid; MFI, median fluorescence intensity; SAP, shrimp alkaline phosphatase; SDS, sodium dodecyl sulfate; SNP, single nucleotide polymorphism.

We have developed and validated a consolidated bead-based genotyping platform, the Bioplex suspension array for simultaneous detection of multiple single nucleotide polymorphisms (SNPs) of the ATP-binding cassette transporters. Genetic polymorphisms have been known to influence therapeutic response and risk of disease pathologies. Genetic screening for therapeutic and diagnostic applications thus holds great promise in clinical management. The allele-specific primer extension (ASPE) reaction was used to assay 22 multiplexed SNPs for eight subjects. Comparison of the microsphere-based ASPE assay results to sequencing results showed complete concordance in genotype assignments. The Bioplex suspension array thus proves to be a reliable, cost-effective and high-throughput technological platform for genotyping. It can be easily adapted to customized SNP panels for specific applications involving large-scale mutation screening of clinically relevant markers.