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Comparative Study
. 2013 Apr;15(2):618-22.
doi: 10.1208/s12248-013-9471-9. Epub 2013 Mar 8.

Direct and rapid genotyping of SLCO1B1 388A>G and 521T>C in human blood specimens using the SmartAmp-2 method

Kenta Yoshida  1 , Junichi Takano, Yuri Ishizu, Alexander Lezhava, Ichiro Ieiri, Kazuya Maeda, Yoshihide Hayashizaki, Yuichi Sugiyama
Affiliations
Comparative Study

Direct and rapid genotyping of SLCO1B1 388A>G and 521T>C in human blood specimens using the SmartAmp-2 method

Kenta Yoshida et al. AAPS J. 2013 Apr.

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.

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Figures

Fig. 1
Fig. 1
Genotyping of SLCO1B1 388A>G and 521T>C by SmartAmp-2. a The primers constructed for the genotyping of SLCO1B1 388A>G and 521T>C SNPs for the SmartAmp-2 assay. b Representative amplification–time profiles generated by the SmartAmp-2 assay in human blood specimens from a, d homozygotes of the WT, b, e heterozygotes, and c, f homozygotes of the mutated type of 388A>G a–c and 521T>C d–f. The primer sets for 388A>G a–c and 521T>C d–f were used in each experiment. Black circles and gray triangles indicate amplifications using the primer set for the WT and MT, respectively. AU arbitrary unit
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