PMID- 31641873 OWN - NLM STAT- MEDLINE DCOM- 20210428 LR - 20210428 IS - 1435-5922 (Electronic) IS - 0944-1174 (Linking) VI - 55 IP - 1 DP - 2020 Jan TI - High-resolution melt analysis enables simple genotyping of complicated polymorphisms of codon 18 rendering the NUDT15 diplotype. PG - 67-77 LID - 10.1007/s00535-019-01638-x [doi] AB - BACKGROUND: The genetic variants of NUDT15 have been verified to induce adverse events (AEs) of thiopurines. Codon 139 variants are frequently observed in Asians, while multiple variants are seen in codon 18 which also cause AEs including the European ancestry. The purpose of this study is to establish a technique capable of the simple genotyping of NUDT15 codon 18 and to evaluate its efficacy. METHODS: A high-resolution melt (HRM) technique is performed to simply determine genotypes. The accuracy of HRM analysis was evaluated with DNAs from 1245 Japanese patients with inflammatory bowel diseases. Subsequently, another group of 572 patients was analyzed to verify the method. The diplotypes and the frequency of their AEs were estimated on the basis of codon 18 and 139 genotypes. RESULTS: The HRM analysis enabled the correct identification of the three main genotypes, ref/ref, ref/ins, and ref/V18I, in 1236 of 1241 cases. All rare genotypes including ref/del were identified as the impossible-to-determine group, the proper diagnosis rate was 99.6%. In the verification test using other samples, the diagnosis rate was 99.7%. By estimating diplotypes using both codon 18 and 139 genotypes, 2.74% and 2.13% of Japanese patients with Arg/Arg and Arg/Cys of codon 139 have a lower enzymatic activity of NUDT15 and a higher risk for adverse responses than those estimated by codon 139 genotypes alone. CONCLUSIONS: Our study showed that HRM method enables simple genotyping of complicated codon 18 variants essential to haplotype estimation of the NUDT15. FAU - Kakuta, Yoichi AU - Kakuta Y AUID- ORCID: 0000-0002-7042-009X AD - Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, 980-8574, Japan. ykakuta@med.tohoku.ac.jp. FAU - Izumiyama, Yasuhiro AU - Izumiyama Y AD - Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, 980-8574, Japan. FAU - Okamoto, Daisuke AU - Okamoto D AD - Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, 980-8574, Japan. FAU - Nakano, Takeru AU - Nakano T AD - Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, 980-8574, Japan. FAU - Ichikawa, Ryo AU - Ichikawa R AD - Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, 980-8574, Japan. FAU - Naito, Takeo AU - Naito T AD - Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, 980-8574, Japan. FAU - Moroi, Rintaro AU - Moroi R AD - Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, 980-8574, Japan. FAU - Kuroha, Masatake AU - Kuroha M AD - Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, 980-8574, Japan. FAU - Kanazawa, Yoshitake AU - Kanazawa Y AD - Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, 980-8574, Japan. FAU - Kimura, Tomoya AU - Kimura T AD - Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, 980-8574, Japan. FAU - Shiga, Hisashi AU - Shiga H AD - Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, 980-8574, Japan. FAU - Kudo, Hisaaki AU - Kudo H AD - Department of Biobank Life Science, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Japan. FAU - Minegishi, Naoko AU - Minegishi N AD - Department of Biobank Life Science, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Japan. FAU - Kawai, Yosuke AU - Kawai Y AD - Genome Medical Science Project, National Center for Global Health and Medicine, Tokyo, Japan. FAU - Tokunaga, Katsushi AU - Tokunaga K AD - Genome Medical Science Project, National Center for Global Health and Medicine, Tokyo, Japan. FAU - Nagasaki, Masao AU - Nagasaki M AD - Human Biosciences Unit for the Top Global Course Center for the Promotion of Interdisciplinary Education and Research (CPIER), Kyoto University, Kyoto, Japan. FAU - Kinouchi, Yoshitaka AU - Kinouchi Y AD - Institute for Excellence in Higher Education, Student Health Care Center, Tohoku University, Sendai, Japan. FAU - Suzuki, Yasuo AU - Suzuki Y AD - Inflammatory Bowel Disease Center, Toho University Sakura Medical Center, Sakura, Japan. FAU - Masasmune, Atsushi AU - Masasmune A AD - Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aoba, Sendai, 980-8574, Japan. CN - MENDEL study group LA - eng GR - 18kk0305002/Japan Agency for Medical Research and Development/ GR - 19ek0410056/Japan Agency for Medical Research and Development/ GR - 16km0405205h0101/Japan Agency for Medical Research and Development/ PT - Journal Article DEP - 20191022 PL - Japan TA - J Gastroenterol JT - Journal of gastroenterology JID - 9430794 RN - 0 (Genetic Markers) RN - 0 (Immunosuppressive Agents) RN - E7WED276I5 (Mercaptopurine) RN - EC 2.6.1.- (NUDT15 protein, human) RN - EC 3.6.1.- (Pyrophosphatases) SB - IM EIN - J Gastroenterol. 2019 Nov 25;:. PMID: 31768800 MH - Female MH - Genetic Markers MH - Genotyping Techniques/*methods MH - Humans MH - Immunosuppressive Agents/*adverse effects/therapeutic use MH - Inflammatory Bowel Diseases/drug therapy/*genetics/metabolism MH - Male MH - Mercaptopurine/*adverse effects/therapeutic use MH - *Pharmacogenomic Variants MH - Pyrophosphatases/*genetics/metabolism OTO - NOTNLM OT - Inflammatory bowel disease OT - NUDT15 OT - Pharmacogenetics OT - Thiopurines FIR - Takagawa, Tetsuya IR - Takagawa T FIR - Nakamura, Shiro IR - Nakamura S FIR - Ikeya, Kentaro IR - Ikeya K FIR - Hanai, Hiroyuki IR - Hanai H FIR - Sakuraba, Hirotake IR - Sakuraba H FIR - Nishida, Atsushi IR - Nishida A FIR - Andoh, Akira IR - Andoh A FIR - Nakagawa, Shoko IR - Nakagawa S FIR - Sasaki, Makoto IR - Sasaki M FIR - Miura, Miki IR - Miura M FIR - Hisamatsu, Tadakazu IR - Hisamatsu T FIR - Toyonaga, Takahiko IR - Toyonaga T FIR - Kobayashi, Taku IR - Kobayashi T FIR - Onodera, Kei IR - Onodera K FIR - Nakase, Hiroshi IR - Nakase H FIR - Shinozaki, Masaru IR - Shinozaki M FIR - Ishiguro, Yoh IR - Ishiguro Y FIR - Mizuno, Shinta IR - Mizuno S FIR - Naganuma, Makoto IR - Naganuma M FIR - Takahara, Masahiro IR - Takahara M FIR - Hiraoka, Sakiko IR - Hiraoka S FIR - Yanai, Shunichi IR - Yanai S FIR - Matsumoto, Takayuki IR - Matsumoto T FIR - Hokari, Ryota IR - Hokari R FIR - Nakagawa, Tomoo IR - Nakagawa T FIR - Araki, Hiroshi IR - Araki H FIR - Motoya, Satoshi IR - Motoya S FIR - Ishihara, Shunji IR - Ishihara S FIR - Oshima, Naoki IR - Oshima N FIR - Katsurada, Takehiko IR - Katsurada T FIR - Sasaki, Yu IR - Sasaki Y FIR - Otsuka, Takafumi IR - Otsuka T FIR - Fujiya, Mikihiro IR - Fujiya M FIR - Onodera, Motoyuki IR - Onodera M FIR - Nagahori, Masakazu IR - Nagahori M FIR - Matsuoka, Katsuyoshi IR - Matsuoka K FIR - Arai, Katsuhiro IR - Arai K FIR - Sato, Yuichiro IR - Sato Y FIR - Noguchi, Mitsunori IR - Noguchi M FIR - Matsuura, Minoru IR - Matsuura M FIR - Ishikawa, Tomoaki IR - Ishikawa T FIR - Nakajima, Hiroki IR - Nakajima H FIR - Terasaki, Hiroshi IR - Terasaki H FIR - Abe, Yukiko IR - Abe Y FIR - Kato, Mai IR - Kato M EDAT- 2019/10/24 06:00 MHDA- 2021/04/29 06:00 CRDT- 2019/10/24 06:00 PHST- 2019/09/12 00:00 [received] PHST- 2019/10/10 00:00 [accepted] PHST- 2019/10/24 06:00 [pubmed] PHST- 2021/04/29 06:00 [medline] PHST- 2019/10/24 06:00 [entrez] AID - 10.1007/s00535-019-01638-x [pii] AID - 10.1007/s00535-019-01638-x [doi] PST - ppublish SO - J Gastroenterol. 2020 Jan;55(1):67-77. doi: 10.1007/s00535-019-01638-x. Epub 2019 Oct 22.