Brief/Technical Note

The AAPS Journal

, Volume 15, Issue 2, pp 618-622

First online:

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

  • Kenta YoshidaAffiliated withLaboratory of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, The University of Tokyo
  • , Junichi TakanoAffiliated withLaboratory of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, The University of Tokyo
  • , Yuri IshizuAffiliated withTechnology Development Unit, Omics Science Center, RIKEN
  • , Alexander LezhavaAffiliated withTechnology Development Unit, Omics Science Center, RIKEN
  • , Ichiro IeiriAffiliated withDepartment of Clinical Pharmacokinetics, Graduate School of Pharmaceutical Sciences, Kyushu University
  • , Kazuya MaedaAffiliated withLaboratory of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, The University of Tokyo
  • , Yoshihide HayashizakiAffiliated withTechnology Development Unit, Omics Science Center, RIKEN
  • , Yuichi SugiyamaAffiliated withSugiyama Laboratory, RIKEN Innovation Center, RIKEN Research Cluster for Innovation Email author 

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