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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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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Acknowledgments
This work was supported partly by a Grant-in-Aid for Scientific Research (S) (24229002) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan.
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Kenta Yoshida and Junichi Takano authors contributed equally to this work.
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Yoshida, K., Takano, J., Ishizu, Y. et al. Direct and Rapid Genotyping of SLCO1B1 388A>G and 521T>C in Human Blood Specimens Using the SmartAmp-2 Method. AAPS J 15, 618–622 (2013). https://doi.org/10.1208/s12248-013-9471-9
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DOI: https://doi.org/10.1208/s12248-013-9471-9