title: Electrochemical detection of Pseudomonas aeruginosa 16S rRNA using a biosensor based on immobilized stem-loop structured probe.
abstract: We have designed an electrochemical DNA biosensor based on stem-loop structured probes for enzymatic detection of Pseudomonas aeruginosa 16S ribosomal RNA (rRNA) in composting degradation. The probe modified with a thiol at its 5' end and a biotin at its 3' end was immobilized on a gold electrode through self-assembly. The stem-loop structured probes were "closed" when target was absent, then the hybridization of the target induced the conformational changes to "open", along with the biotin at its 3' end binding with streptavidin-horseradish peroxidase (HRP), and subsequent quantiﬁcation of the target was detected via electrochemical detecting the enzymatic product in the presence of substrate. Under the optimum experiment conditions, the amperometric current response to HRP-catalyzed reaction was linearly related to the logarithm of the target nucleic acid concentration, ranging from 0.3 and 600 pg/μL, with the detection limit of 0.012 pg/μL. A correlation coefficient of 0.9960 was identified. The 16S rRNA extracted from P. aeruginosa was analyzed by this proposed sensor. The results were in agreement with the reference values deduced from UV spectrometric data. The biosensor was indicative of good precision, stability, sensitivity, and selectivity.
authors: College of Environmental Science and Engineering, Hunan University, Changsha 410082, China. liucan8407@163.com
authors: Can
authors: C
authors: Liu
authors: Guang-Ming
authors: GM
authors: Zeng
authors: Lin
authors: L
authors: Tang
authors: Yi
authors: Y
authors: Zhang
authors: Yuan-Ping
authors: YP
authors: Li
authors: Yuan-Yuan
authors: YY
authors: Liu
authors: Zhen
authors: Z
authors: Li
authors: Meng-Shi
authors: MS
authors: Wu
authors: Jie
authors: J
authors: Luo
mesh_terms: D001673:Biodegradation, Environmental; D015374:Biosensing Techniques; D019921:Biotinylation; D002138:Calibration; D015342:DNA Probes / Q000737:chemistry*; D055664:Electrochemical Techniques; D004566:Electrodes; D004867:Equipment Design; D006017:Glycolipids / Q000378:metabolism; D006735:Horseradish Peroxidase; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization / Q000379:methods*; D056951:Photoelectron Spectroscopy; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D012329:RNA, Bacterial / Q000032:analysis*; D012336:RNA, Ribosomal, 16S / Q000032:analysis*; D012680:Sensitivity and Specificity; D012988:Soil Microbiology; D019809:Streptavidin; D013501:Surface-Active Agents / Q000378:metabolism
chemical_list: D015342:DNA Probes; D006017:Glycolipids; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D013501:Surface-Active Agents; C418382:rhamnolipid; D019809:Streptavidin; D006735:Horseradish Peroxidase