Department of Civil Engineering, National Institute of Technology.
Division of Environmental Engineering, Faculty of Engineering, Hokkaido University.
Microbes Environ. 2022;37(1). doi: 10.1264/jsme2.ME21073.
An endpoint recombination amplification reaction (RPA) assay for assessing the abundance of the gene encoding thiocyanate dehydrogenase (TcDH) in Thiohalobacter has been developed. The RPA reaction was performed at 37°C for 30 min, terminated by the addition of sodium dodecyl sulfate (SDS) solution, and the DNA concentration of the RPA product was fluorometrically measured. The abundance of TcDH in 22 activated sludge samples and 7 thiocyanate-degrading enrichment cultures ranged between 2.5×10 and 1.5×10 copies μL, showing a linear relationship (R=0.83) with those measured using a conventional quantitative PCR assay.
已经开发出一种用于评估硫氰酸盐脱氢酶（TcDH）基因丰度的终点重组扩增反应（RPA）测定法。RPA 反应在 37°C 下进行 30 分钟，通过加入十二烷基硫酸钠（SDS）溶液终止，并通过荧光法测量 RPA 产物的 DNA 浓度。在 22 个活性污泥样品和 7 个硫氰酸盐降解富集培养物中 TcDH 的丰度在 2.5×10 和 1.5×10 拷贝μL 之间，与使用常规定量 PCR 测定法测量的丰度呈线性关系（R=0.83）。
