イマイ　カズオ   IMAI Kazuo   今井　一男    所属   埼玉医科大学  医学部 臨床検査医学（中央検査部）    職種   専任講師
論文種別	学術雑誌(原著)
言語種別	英語
査読の有無	査読なし
表題	A novel peptide nucleic acid- and loop-mediated isothermal amplification assay for the detection of mutations in the 23S rRNA gene of Treponema pallidum.
掲載誌名	正式名：Journal of medical microbiology
巻・号・頁	69(12),1339-1345頁
著者・共著者	Norihito Tarumoto,Kazuo Imai,Shu-Ichi Nakayama,Ichiro Itoda,Jun Sakai,Takashi Murakami,Shigefumi Maesaki,Satoshi Hayakawa,Makoto Ohnishi,Takuya Maeda
発行年月	2020/12
概要	Introduction. Macrolides could be a potential alternative treatment for Treponema pallidum infections in patients; however, macrolide-resistant T. pallidum is spreading rapidly worldwide.Hypothesis/Gap Statement. There are presently no alternatives to serological tests for syphilis that can be used to evaluate therapeutic effects due to the fact that T. pallidum cannot be cultured in vitro.Aim. In this study, we constructed a method for rapidly identifying T. pallidum and confirming macrolide resistance by using loop-mediated isothermal amplification (LAMP) with peptide nucleic acids (PNAs).Methodology. A set of LAMP primers was designed to span nucleotide positions 2058 and 2059 in 23S rRNA. A PNA clamping probe was also designed to be complementary to the wild-type sequence (A2058/A2059) and positioned to interfere with both the annealing of the 3' end of the backward inner primer and the concomitant extension. Prior to the LAMP assay, swab samples from suspected syphilitic lesions were boiled for DNA extraction.Results. The assay had an equivalent detection limit of 1.0×101 copies/reaction and showed specificity against 38 pathogens. In the presence of a 4 µM PNA probe, LAMP
DOI	10.1099/jmm.0.001275
PMID	33180016