title: Determination for Enterobacter cloacae based on a europium ternary complex labeled DNA probe.
abstract: The fast detection and accurate diagnosis of the prevalent pathogenic bacteria is very important for the treatment of disease. Nowadays, fluorescence techniques are important tools for diagnosis. A two-probe tandem DNA hybridization assay was designed for the detection of Enterobacter cloacae based on time-resolved fluorescence. In this work, the authors synthesized a novel europium ternary complex Eu(TTA)(3)(5-NH(2)-phen) with intense luminescence, high fluorescence quantum yield and long lifetime before. We developed a method based on this europium complex for the specific detection of original extracted DNA from E. cloacae. In the hybridization assay format, the reporter probe was labeled with Eu(TTA)(3)(5-NH(2)-phen) on the 5'-terminus, and the capture probe capture probe was covalent immobilized on the surface of the glutaraldehyde treated glass slides. The original extracted DNA of samples was directly used without any DNA purification and amplification. The detection was conducted by monitoring the fluorescence intensity from the glass surface after DNA hybridization. The detection limit of the DNA was 5×10(-10) mol L(-1). The results of the present work proved that this new approach was easy to operate with high sensitivity and specificity. It could be conducted as a powerful tool for the detection of pathogen microorganisms in the environment.
authors: Engineering department, Rainbow Department Store CO., LTD., Shenzhen 518026, PR China.
authors: Hui
authors: H
authors: He
authors: Cheng-Gang
authors: CG
authors: Niu
authors: Guang-Ming
authors: GM
authors: Zeng
authors: Min
authors: M
authors: Ruan
authors: Pin-Zhu
authors: PZ
authors: Qin
authors: Jing
authors: J
authors: Liu
mesh_terms: D004247:DNA / Q000737:chemistry*; D015342:DNA Probes / Q000737:chemistry*; D016972:Enterobacter cloacae / Q000378:metabolism*; D005063:Europium / Q000737:chemistry*; D005898:Glass; D005976:Glutaral / Q000737:chemistry; D008956:Models, Chemical; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000737:chemistry; D012037:Refuse Disposal; D015203:Reproducibility of Results; D012722:Sewage; D013050:Spectrometry, Fluorescence / Q000379:methods; D013997:Time Factors
chemical_list: D015342:DNA Probes; D009841:Oligonucleotides; D012722:Sewage; D005063:Europium; D004247:DNA; D005976:Glutaral