Abstract
We developed a method for single cell analysis with simultaneous detection of a functional gene and rRNA by using two-pass tyramide signal amplification-fluorescence in situ hybridization (TSA-FISH) for identification of denitrifying bacteria in activated sludge from a night soil treatment plant. The new method enabled simultaneous detection of the nirK gene and rRNA from pure cultures at a high detection rate. Microbial communities in the sludge were identified using cloning analysis based on rRNA genes and the nirK gene. On the basis of the results of microbial community analysis, we designed probes targeting rRNA and the nirK gene. Simultaneous detection of nirK gene and rRNA identified Hyphomicrobium as a denitrifier in the sludge.
