| Title: | Application of quantitative real-time PCR for quantification of Rhodococcus sp. EH831 in a polyurethane biofilter |
| Address: | "Department of Environmental Science and Engineering, Ewha Womans University, Seoul, Republic of Korea" |
| ISSN/ISBN: | 0254-8704 (Print) 0254-8704 (Linking) |
| Abstract: | "Rhodococcus sp. EH831 is a microbial species that can degrade volatile organic compounds. We optimized a method for monitoring quantitative real-time PCR (qRT-PCR) of EH831 that was incorporated into a polyurethane (PU) biofilter. When the genomic DNA of EH831 was directly extracted from a PU sample with immobilized EH831, the recovery efficiency was very low due to DNA absorption into the PU. DNA amplification during PCR was also inhibited by PU impurities. Therefore, a pre-treatment step was necessary. We successfully recovered cells from the PU by squeezing the matrix, adding sterilized water, and vortexing. The recovery efficiency ranged from 105 to 144%, and there was no statistically significant difference. We designed a novel TaqMan probe for EH831 and demonstrated its high specificity for EH831. The detection range for EH831 was 10(5)-10(11) CFU ml(-1). The method described in this study can be used to investigate the relationship between quantitative analysis of Rhodococcus sp. EH831 and PU biofilter performance" |
| Keywords: | "DNA, Bacterial/chemistry Filtration/instrumentation Polymerase Chain Reaction/*methods *Polyurethanes RNA, Ribosomal, 16S/chemistry Rhodococcus/*genetics/growth & development/metabolism;" |
| Notes: | "MedlineLee, E H Cho, K S Ryu, H W eng Evaluation Study Research Support, Non-U.S. Gov't India 2010/02/02 J Environ Biol. 2009 Jan; 30(1):155-9" |
