Duplex pyrosequencing assay of the 388A>G and 521T>C SLCO1B1 polymorphisms in three Asian populations

Abstract

BACKGROUND: We developed a method for detecting important SLCO1B1 polymorphisms and compared the haplotype frequencies in 3 Asian populations. METHODS: We designed a duplex pyrosequencing assay to detect simultaneously the 388A>G and 521T>C variants of SLCO1B1; this method can identify SLCO1B1*1b, SLCO1B1*5, and SLCO1B1*15. The method was validated by direct sequencing of 96 Korean subjects. In addition, duplex genotyping and the monoplex method were compared and validated with 469 Korean subjects. To characterize the haplotype frequencies based on the 2 polymorphisms, we genotyped 106 Chinese and 104 Vietnamese subjects, as well as Korean subjects, using the new method. RESULTS: The results showed 100% concordance among the monoplex and duplex pyrosequencing assays and direct sequencing method. The allele frequencies were similar in the 3 Asian populations: the most common allele was SLCO1B1*1b, while SLCO1B1*5 was rare or absent. The frequencies of functional SLCO1B1*15 alleles differed statistically between Chinese (8.2%) and Korean (14.0%) and Vietnamese (16.3%) (p<0.05, chi(2)-test). CONCLUSIONS: The duplex pyrosequencing assay appears to be an accurate, rapid, and cost-effective genotyping method to detect major SLCO1B1 important alleles in Asian populations.