文章基本信息

标题：Genotyping of Thiopurine Methyltransferase Using PyrosequencingTM
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作者：Yuko Okada ; Katsunori Nakamura ; Masanori Wada 等
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：2005
卷号：28
期号：4
页码：677-681
DOI：10.1248/bpb.28.677
出版社：The Pharmaceutical Society of Japan
摘要：Thiopurine methyltransferase (TPMT) metabolizes thiopurine drugs which are used in the treatment of leukemia and some autoimmune diseases. Previously, 11 mutant alleles of TPMT gene ( TPMT*1S , *2 , *3A , *3B , *3C , *3D , *4 , *5 , *6 , *7 , and *8 ) have been reported. These mutant alleles may cause life-threatening toxicity in patients exposed to thiopurine drugs, 6-mercaptopurine and azathioprine. We have developed a rapid and accurate protocol for TPMT genotype determination using Pyrosequencing^TM technology in 96 Japanese subjects. Five fragments of the TPMT gene (exon 4, 5, 7, 8, 10) were amplified by PCR, and the 10 single-nucleotide polymorphisms (SNPs) for TPMT*1S , *2 , *3A , *3B , *3C , *3D , *4 , *5 , *6 , *7 , and *8 were sequenced. The results of this pyrosequencing method corresponded exactly with those of the DNA sequencing method using BigDye terminator chemistry. We have demonstrated that typing of 10 SNPs can be performed within 30 min. Pyrosequencing has a wide application in the large-scale identification of individual TPMT genotypes.
关键词：thiopurine methyltransferase;pyrosequencing;polymorphism;6-mercaptopurine