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	査読あり
表題	Development of a highly resolved loop-mediated isothermal amplification method to detect the N526K ftsl mutation of beta-lactamase-negative ampicillin-resistant Haemophilus influenzae
掲載誌名	正式名：JOURNAL OF MICROBIOLOGICAL METHODS ISSNコード：01677012
出版社	ELSEVIER SCIENCE BV
巻・号・頁	141,108-114頁
著者・共著者	Shinsuke Tamura,Takuya Maeda,Kazuhisa Misawa,Morichika Osa,Takaaki Hamamoto,Atsushi Yuki,Kazuo Imai,Kei Mikita,Kyoko Morichika,Akihiko Kawana,Hiroshi Matsumoto,Shigeaki Nonoyama
発行年月	2017/10
概要	Rapid and easy detection of sequence polymorphisms, including nucleotide point mutations of bacterial pathogens responsible for amino acid substitutions linked to drug resistance, is essential for the proper use of antimicrobial agents. Here, a detection method using loop-mediated amplification (LAMP) combined with amplification refractory mutation system (ARMS) to accurately distinguish a different single nucleotide in the target sequence was established, named ARMS-SNP LAMP. This procedure is capable of species-specific detection of a nucleotide (1578T) in the ftsl gene on Haemophilus influenzae without amplifying the sequence carrying the point mutations (T1578G/A) in beta-lactamase-negative ampicillin resistant (BLNAR) strains. Reactions were performed at 61 degrees C for 45 min. Successful target gene amplifications were detected by measuring real-time turbidity using a turbidimeter and visual detection. The assay had a detection limit of 10.0 pg of genomic DNA per reaction and showed specificity against 52 types of pathogens, whereas amplifications were completely blocked in even 100.0 ng/mu L of genomic DNA with point mutations at T1578G and T1578A. The expected ARMS-SNP LAM
DOI	10.1016/j.mimet.2017.08.008
PMID	28807759