DEVELOPMENT OF THE QUANTITATIVE PCR METHOD FOR Candidatus ACCUMULIBACTER PHOSPHATIS AND ITS APPLICATION TO ACTIVATED SLUDGE
To quantify Candidatus Accumulibacter phosphatis in activated sludge, quantitative PCR method was developed utilizing SYBR GREEN I and a specific primer set targeted on the 16S rDNA of Candidatus Accumulibacter phosphatis. Following optimization of PCR condition, specificity was evaluated based on the melting curve and the sequencing analysis of the PCR products with DNA extracted from activated sludge. Both the melting curve and the sequencing analysis of the PCR product showed that only the target DNA from Candidatus Accumulibacter phosphatis was amplified. Standard curves with a series of tenfold dilution of the DNA from 16S rDNA fragment of Candidatus Accumulibacter phosphatis gave R2 values greater than 0.999. The minimum detection limit was.1.0×103 copies per reaction. The amount of Candidatus Accumulibacter phosphatis in five laboratory-scale and ten full-scale activated sludge samples were quantified both by the quantitative PCR method and by the FISH method. The quantification results by these two methods agreed satisfactorily, with an R2 value of 0.6871 showing a statistically significant correlation (p<0.001). Thus, we developed a rapid quantification method by using quantitative PCR for the quantification of Candidatus Accumulibacter phosphatis in activated sludge.
Quantitative PCR SYBR GREEN I Candidatus Accumulibacter phosphatis Activated Sludge
Fukushima Naoki Uda Motoharu Onuki Hiroyasu Satoh Takashi Mino
Institute of Environmental Studies, Graduate School of Frontier Sciences, The University of Tokyo, 5 Integrated Research System for Sustainability Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-
国际会议
第六届中日水环境暨NSFC-JST重大国际合作项目成果交流会
青岛
英文
59-63
2006-10-19（万方平台首次上网日期，不代表论文的发表时间）