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Multiplexed genotyping of ABC transporter polymorphisms with the Bioplex suspension array

Abstract

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.

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

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Correspondence to Edmund Jon Deoon Lee.

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Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/2.0 ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Koo, S.H., Ong, T.C., Chong, K.T. et al. Multiplexed genotyping of ABC transporter polymorphisms with the Bioplex suspension array. Biol. Proced. Online 9, 27–42 (2007). https://doi.org/10.1251/bpo131

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  • DOI: https://doi.org/10.1251/bpo131

Indexing terms

  • Genotype
  • Microspheres
  • Polymorphism, Genetic