The AAPS Journal

, Volume 15, Issue 2, pp 618–622

Direct and Rapid Genotyping of SLCO1B1 388A>G and 521T>C in Human Blood Specimens Using the SmartAmp-2 Method

  • Kenta Yoshida
  • Junichi Takano
  • Yuri Ishizu
  • Alexander Lezhava
  • Ichiro Ieiri
  • Kazuya Maeda
  • Yoshihide Hayashizaki
  • Yuichi Sugiyama
Brief/Technical Note

Abstract

Organic anion-transporting polypeptide (OATP) 1B1, encoded by the solute carrier organic anion transporter family member 1B1 (SLCO1B1) gene, mediates the active uptake of various organic anions into hepatocytes and determines their hepatic clearances as the first step in the detoxification pathway. Previous reports indicated that alterations in its function by drug–drug interactions or genetic polymorphisms affect the pharmacokinetics of the substrate drugs. In the present study, we developed a method to genotype SLCO1B1 388A>G (rs2306283) and 521>C (rs4149056), which significantly affect the clinical pharmacokinetics and subsequent side effects such as myopathy caused by statins, OATP1B1 substrates in humans. We used a small aliquot of blood and the isothermal Smart Amplification Process version 2 (SmartAmp-2), which could complete the genotyping of 388A>G and 521T>C within 60 min. The genotypes of 101 genomic DNA samples and blood samples assessed by SmartAmp-2 matched perfectly to those determined previously by the conventional PCR-SSCP method. The SmartAmp-2 method enables the rapid identification of the 388A>G and 521T>C genotypes, saving time and effort in the genomic DNA preparation in clinical practice. This method will be useful for evaluating and predicting altered pharmacological and toxicological effects of substrate drugs caused by SLCO1B1 polymorphisms.

KEY WORDS

membrane transporters pharmacogenetics single nucleotide polymorphisms 

Abbreviations

AmC7-Q

3′-Amino modifier C7

AUC

Area under the concentration–time curve

HMG-CoA

Hydroxymethylglutaryl-CoA

MT

Mutated type

OATP1B1

Organic anion-transporting polypeptide 1B1

RFLP

Restriction fragment length polymorphism

SLCO1B1

Solute carrier organic anion transporter family member 1B1

SmartAmp-2

Smart Amplification Process version 2

SNP

Single nucleotide polymorphism

SSCP

Single-strand conformation polymorphism

WT

Wild type

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Copyright information

© American Association of Pharmaceutical Scientists 2013

Authors and Affiliations

  • Kenta Yoshida
    • 1
  • Junichi Takano
    • 1
  • Yuri Ishizu
    • 2
  • Alexander Lezhava
    • 2
  • Ichiro Ieiri
    • 3
  • Kazuya Maeda
    • 1
  • Yoshihide Hayashizaki
    • 2
  • Yuichi Sugiyama
    • 4
  1. 1.Laboratory of Molecular Pharmacokinetics, Graduate School of Pharmaceutical SciencesThe University of TokyoTokyoJapan
  2. 2.Technology Development Unit, Omics Science CenterRIKENYokohamaJapan
  3. 3.Department of Clinical Pharmacokinetics, Graduate School of Pharmaceutical SciencesKyushu UniversityFukuokaJapan
  4. 4.Sugiyama Laboratory, RIKEN Innovation CenterRIKEN Research Cluster for InnovationYokohamaJapan

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