LOOP-MEDIATED ISOTHERMAL AMPLIFICATION FOR QUANTIFICATION OF DEHALOCOCCOIDES AND ITS APPLICATION TO PRACTICAL ANAEROBIC BIOSTIMULATION OF GROUNDWATER CONTAMINATED WITH CHLORINATED ETHENES

Abstract

 This study developed novel loop-mediated isothermal amplification (LAMP) methods for specific and quantitative monitoring of Dehalococcoides spp. using 16S rRNA gene and dehalogenase genes bvcA and vcrA as the target genes. The quantification limit for the developed LAMP methods was 1 × 10³ copies per microliter-DNA sample, and the quantification results of the developed methods correlated positively with those of real-time PCR, which has been commonly applied for monitoring Dehalococcoides spp. Application of the developed LAMP methods to the practical anaerobic biostimulation of groundwater contaminated with chlorinated ethenes, in addition to the measurements of conventional physical and chemical indicators, demonstrated that the penetration of injected nutritional materials, change to a strong anaerobic condition, increase of Dehalococcoides spp., decomposition of chlorinated ethenes occurred in sequence. These results indicated that the LAMP methods developed here enable simple and reliable monitoring of Dehalococcoides spp. and are helpful in the management of in situ bioremediation.