Development of a loop-mediated isothermal amplification method for the rapid detection of the dioxin-degrading bacterium Ochrobactrum anthropi in soil

J Environ Manage. 2015 Sep 1:160:263-70. doi: 10.1016/j.jenvman.2015.06.027. Epub 2015 Jul 3.

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.

Keywords: Biomonitoring; Dioxin; LAMP; Loop-mediated isothermal amplification; Ochrobactrum anthropi BD-1.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Primers
  • DNA, Bacterial / analysis
  • Dioxins / chemistry*
  • Humans
  • Nucleic Acid Amplification Techniques / methods*
  • Ochrobactrum anthropi / genetics*
  • Reproducibility of Results
  • Soil Microbiology*

Substances

  • DNA Primers
  • DNA, Bacterial
  • Dioxins