title: Detection of bacteria carrying the stx2 gene by in situ loop-mediated isothermal amplification.
abstract: A new in situ DNA amplification technique for microscopic detection of bacteria carrying a specific gene is described. Loop-mediated isothermal amplification (LAMP) was used to detect stxA(2) in Escherichia coli O157:H7 cells. The mild permeabilization conditions and low isothermal temperature used in the in situ LAMP method caused less cell damage than in situ PCR. It allowed use of fluorescent antibody labeling in the bacterial mixture after the DNA amplification for identification of E. coli O157:H7 cells with an stxA(2) gene. Higher-contrast images were obtained with this method than with in situ PCR.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
authors.forename: Fumito
authors.initials: F
authors.lastname: Maruyama
authors.forename: Takehiko
authors.initials: T
authors.lastname: Kenzaka
authors.forename: Nobuyasu
authors.initials: N
authors.lastname: Yamaguchi
authors.forename: Katsuji
authors.initials: K
authors.lastname: Tani
authors.forename: Masao
authors.initials: M
authors.lastname: Nasu
mesh_terms: D019453:Escherichia coli O157 / Q000235:genetics*; D005784:Gene Amplification; D017404:In Situ Hybridization, Fluorescence; D016133:Polymerase Chain Reaction; D022641:Shiga Toxin 2 / Q000235:genetics*
chemical_list: D022641:Shiga Toxin 2