Shimizu Makiko, Mizugaki Ami, Koibuchi Natsumi, Sango Haruna, Uenuma Yumi, Yamazaki Hiroshi
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Tokyo, Japan.
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Tokyo, Japan.
Drug Metab Pharmacokinet. 2021 Dec;41:100420. doi: 10.1016/j.dmpk.2021.100420. Epub 2021 Oct 8.
Increasing numbers of single-nucleotide substitutions of the human flavin-containing monooxygenase 3 (FMO3) gene are being recorded in mega-databases. Phenotype-gene analyses revealed impaired FMO3 variants associated with the metabolic disorder trimethylaminuria. Here, a series of reliable FMO3 genotyping confirmation methods was assembled and developed for 45 impaired FMO3 variants, mainly found in Japanese populations, using singleplex or duplex polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) methods and singleplex, duplex, or tetraplex allele-specific PCR methods. Nine PCR-RFLP procedures with single restriction enzymes and fourteen duplex PCR-RFLP procedures (for p.Trp41Ter and p.Thr329Ala, p.Met66Val and p.Leu163Pro, p.Pro70Leu and p.Glu308Gly, p.Asn114Ser and p.Ser195Leu, p.Glu158Lys and p.Ile441Thr, p.Cys197Ter and p.Trp388Ter, p.Arg205Cys and p.Val257Met, p.Arg205His and p.Cys397Ser, p.Met211ArgfsTer10 and p.Arg492Trp, p.Arg223Gln and p.Leu473Pro, p.Met260Val and p.Thr488Ala, p.Tyr269His and p.Ala311Pro, p.Ser310Leu and p.Gly376Glu, and p.Gln470Ter and p.Arg500Ter) were newly established along with eight singleplex (for p.Pro153GlnfsTer14, p.Gly191Cys, p.Pro248Thr, p.Ile486Met, and p.Pro496Ser, among others), one duplex (p.Ile199Ser and p.Asp286Tyr), and one tetraplex (p.Ile7Thr, p.Val58Ile, p.Thr201Lys, and p.Gly421Val) allele-specific PCR systems. This series of systems should facilitate the easy detection in a clinical setting of FMO3 variants in Japanese subjects susceptible to low drug clearances or drug reactions possibly caused by impaired FMO3 function.
在大型数据库中记录到的人类含黄素单加氧酶3（FMO3）基因的单核苷酸替换数量不断增加。表型-基因分析显示，FMO3变体受损与代谢紊乱三甲胺尿症相关。在此，针对主要在日本人群中发现的45种受损FMO3变体，使用单重或双重聚合酶链反应（PCR）-限制性片段长度多态性（RFLP）方法以及单重、双重或四重等位基因特异性PCR方法，组装并开发了一系列可靠的FMO3基因分型确认方法。新建立了9种使用单一限制性内切酶的PCR-RFLP程序和14种双重PCR-RFLP程序（针对p.Trp41Ter和p.Thr329Ala、p.Met66Val和p.Leu163Pro、p.Pro70Leu和p.Glu308Gly、p.Asn114Ser和p.Ser195Leu、p.Glu158Lys和p.Ile441Thr、p.Cys197Ter和p.Trp388Ter、p.Arg205Cys和p.Val257Met、p.Arg205His和p.Cys397Ser、p.Met211ArgfsTer10和p.Arg492Trp、p.Arg223Gln和p.Leu473Pro、p.Met260Val和p.Thr488Ala、p.Tyr269His和p.Ala311Pro、p.Ser310Leu和p.Gly376Glu，以及p.Gln470Ter和p.Arg500Ter），同时还建立了8种单重（针对p.Pro153GlnfsTer14、p.Gly191Cys、p.Pro248Thr、p.Ile486Met和p.Pro496Ser等）、1种双重（p.Ile199Ser和p.Asp286Tyr）和1种四重（p.Ile7Thr、p.Val58Ile、p.Thr201Lys和p.Gly421Val）等位基因特异性PCR系统。这一系列系统应有助于在临床环境中轻松检测日本受试者中可能因FMO3功能受损导致药物清除率低或药物反应的FMO3变体。
