title: [Development and evaluation of novel loop-mediated isothermal amplification for rapid detection of bla(IMP-1) and bla(VIM-2) genes].
abstract: Loop-mediated isothermal amplification (LAMP) amplifies a target gene with high specificity and rapidity under isothermal conditions. LAMP assays were developed for the rapid detection of metallo-beta-lactamase (MBL) genes such as bla(IMP-1)) and bla(VIM-2). We initially designed specific primers to detect MBL genes for LAMP assays and evaluated the specificity and sensitivity of these assays. LAMP assays amplified MBL genes under a constant temperature of 63 degrees C within 1 hour, and were compared to PCR in MBL-producing strains. The results of MBL genes typing by LAMP assays agree completely with PCR results. The lower detection limits of bla(IMP-1)- and bla(VIM-2)-LAMP assays using real-time turbidimeters were 30cfu/test and 3cfu/test. After amplification, products were directly observed by the naked eye with a fluorescent detection reagent. In conclusion, LAMP assays are convenient, rapid, and fully feasible for detecting MBL genes in ordinary clinical microbiology laboratories without special apparatus.
authors.affiliation: Biochemical Research Laboratory, Eiken Chemical Co., Ltd.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Kojima
authors.forename: Naohiro
authors.initials: N
authors.lastname: Shibata
authors.forename: Masanari
authors.initials: M
authors.lastname: Ikedo
authors.forename: Yoshichika
authors.initials: Y
authors.lastname: Arakawa
mesh_terms: D005784:Gene Amplification; D012680:Sensitivity and Specificity; D001618:beta-Lactamases / Q000235:genetics*
chemical_list: D001618:beta-Lactamases