title: Development of a loop-mediated isothermal amplification method for the rapid detection of the dioxin-degrading bacterium Ochrobactrum anthropi in soil.
abstract: In this study, loop-mediated isothermal amplification (LAMP) and real-time LAMP assays were developed to detect the dioxin-degrading bacterium Ochrobactrum anthropi strain BD-1 in soil. Four primers were designed to use ITS gene amplification for the strain O. anthropi BD-1. The real-time LAMP assay was found to accomplish the reaction by 1 pg of genomic DNA load when used for nucleic acid amplification. This assay was then applied to detect O. anthropi BD-1 in eight soil samples collected from a dioxin-contaminated site. The results demonstrated that these newly developed LAMP and real-time LAMP assays will not only be useful and efficient tools for detecting the target gene, but also be used as molecular tools for monitoring the growth of dioxin-degrading O. anthropi in the soil. This is the first report to demonstrate the use of LAMP assays to monitor the presence of O. anthropi in dioxin-contaminated soil. The application of this method should improve the biomonitoring of dioxin contamination.
authors.affiliation: Department of Safety, Health, and Environmental Engineering, Ming Chi University of Technology, New Taipei City 24301, Taiwan. Electronic address: hjchen@mail.mcut.edu.tw.
authors.forename: Hsi-Jien
authors.initials: HJ
authors.lastname: Chen
authors.affiliation: Graduate Institute of Pharmaceutical Chemistry, College of Pharmacy, China Medical University, Taichung 40402, Taiwan.
authors.forename: Meng-Shiou
authors.initials: MS
authors.lastname: Lee
authors.affiliation: Department of Safety, Health, and Environmental Engineering, Ming Chi University of Technology, New Taipei City 24301, Taiwan.
authors.forename: Jun-Yu
authors.initials: JY
authors.lastname: Lai
authors.affiliation: Graduate Institute of Biotechnology, College of Agriculture and Natural Resources, National Chung Hsing University, Taichung 40402, Taiwan
authors.forename: Guan-Hua
authors.initials: GH
authors.lastname: Lai
mesh_terms: D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D004147:Dioxins / Q000737:chemistry*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D020621:Ochrobactrum anthropi / Q000235:genetics*; D015203:Reproducibility of Results; D012988:Soil Microbiology
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D004147:Dioxins
keywords: Biomonitoring; Dioxin; LAMP; Loop-mediated isothermal amplification; Ochrobactrum anthropi BD-1