Ralstonia eutrophaKT-1 is a bacterium able to degrade trichloroethylene (TCE) by its phenol or toluene-degrading enzyme. Quantitative detection of strain KT-1 was examined with a real time PCR apparatus, in order to monitor the strain released into a field for TCE degradation. A 800 base-DNA fragment, PCR-synthesized by using a primer pair designed based on a repetitive extragenic palindromic (REP) sequence, was used as a target DNA for the detection. A primer pair with hybridization probes, designed from the sequence of the PCR-synthesized fragment, successfully detected strain KT-1 and detection limit was 20 cell-DNA/PCR-tube. This developed method was applied to the detection of strain KT-1 injected to ground water of a TCE contaminated site in Chiba prefecture. Strain KT-1 in the groundwater was monitored for approximately two months after the injection. The monitoring of strain KT-1 was successfully attained until the detection limit, showing the effectiveness of the quantitative PCR.
