Development of pyrosequencing method for detection of ABCG2 polymorphisms

Abstract

Abstract: AIM: To establish a pyrosequencing based method for detection ABCG2 34G＞A and 421C＞A polymorphisms and to determine the frequency of these polymorphisms in healthy Chinese. METHODS: After preparation of gDNA from blood of 200 subjects, the target fragments were amplified by PCR, polymorphisms were detected on PyroMark ID by pyrosequencing technology. The reliability of pyrosequencing methods were validated by repeat tests and Sanger sequencing. RESULTS: We established a new pyrosequencing method to detect the ABCG2 34G＞A and 421C＞A polymorphisms polymorphisms in healthy Chinese. The detection rate and repetition rate were both 100%. The frequencies of ABCG2 34G and 34A alleles were 79.5% and 20.5%, respectively. The allele frequencies of ABCG2 421C and 421A were 72.7% and 27.8%, respectively. Genotype frequencies match the Hardy-Weinberg equilibrium. CONCLUSION: These pyrosequencing assays to detect ABCG2 polymorphisms are proved to be a rapid, accurate and high-throughput alternative to conventional methods, and it can be a preferred option in research and clinical application.

Key words: Pharmacology, Pyrosequencing, ABCG2, BCRP, SNPs

CLC Number:

R968

WAN Zi-rui, XIE Hai-tang, YU Jing, LIU Ying-zi, ZENG Fei-yue, WANG Guo. Development of pyrosequencing method for detection of ABCG2 polymorphisms[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(7): 779-784.

References

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