title: Determination of ammonia-oxidizing bacteria based on time-resolved fluorescence and tandem probe.
abstract: To study the ecology of ammonia-oxidizing bacteria (AOB), quantitative techniques are essential. In this report, the authors introduced an innovative method based on time-resolved fluorescence to quantify AOB as a representative of the major functional microorganisms in sewage water treatment. A bifunctional europium complex with the characteristics of long lifetime and intense luminescence was used as labels in the experiments. In the detection, the capture probe and dye-labeled reporter probe could form a two-probe tandem with the target sequence, and the determination of target DNA was done by monitoring the time-resolved fluorescence signals of europium complex-labeled reporter probe left on the glass slide surface. The experiment conditions consisting of concentration of capture probe, hybridization temperature, hybridization time and washing time were optimized. This method presents satisfactory specificity to common bacteria such as Nitrobacter winogradskyi, Escherichia coli and Paenibacillus polymyxa. The detection limit was 3.65 × 10(-11)mol L(-1). This detection system enables us to rapidly and sensitively analyze the microbial population variety in sewage water treatment.
authors: College of Environmental Science and Engineering, Key Laboratory of Environmental Biology and Pollution Control, Ministry of Education, Hunan University, Changsha 410082, China.
authors: Xiaoxiao
authors: X
authors: Lv
authors: Zhizhang
authors: Z
authors: Li
authors: Chenggang
authors: C
authors: Niu
authors: Min
authors: M
authors: Ruan
authors: Dawei
authors: D
authors: Hang
authors: Guangming
authors: G
authors: Zeng
mesh_terms: D000641:Ammonia / Q000378:metabolism*; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004783:Environmental Microbiology; D015200:Fluoroimmunoassay / Q000379:methods*; D010084:Oxidation-Reduction; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial; D000641:Ammonia