Electrochemical detection of Pseudomonas aeruginosa 16S rRNA using a biosensor based on immobilized stem-loop structured probe
Electrochemical detection of Pseudomonas aeruginosa 16S rRNA using a biosensor based on immobilized stem-loop structured probe
复制标题
基于固定化茎环结构探针的生物传感器电化学检测铜绿假单胞菌16S rRNA
DOI：
10.1016/j.enzmictec.2011.06.011
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发表时间：
2011-08-10
影响因子：
3.4
通讯作者：
Luo, Jie
中科院分区：
文献类型：
--
作者：
Liu, Can;Zeng, Guang-Ming;Luo, Jie
我们设计了一种基于茎环结构探针的电化学DNA生物传感器，用于检测堆肥降解过程中铜绿假单胞菌16 S核糖体RNA（rRNA）。通过自组装将5'端巯基、3'端生物素修饰的探针固定在金电极上。当靶不存在时，茎环结构探针是“闭合的”，然后靶的杂交诱导构象变化为“开放的”，沿着，其3'端的生物素与链霉亲和素-辣根过氧化物酶（HRP）结合，随后在底物存在下通过电化学检测酶促产物来检测靶的定量。在最佳实验条件下，HRP催化反应的安培电流响应与靶核酸浓度的对数呈线性关系，线性范围为0.3 ~ 600 pg/μ L，检出限为0.012 pg/μ L。相关系数为0.9960。利用该传感器对从铜绿假单胞菌中提取的16 S rRNA进行了分析。测定结果与紫外光谱数据的参考值一致。该生物传感器具有良好的精密度、稳定性、灵敏度和选择性。(C)2011 Elsevier Inc. All rights reserved.
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/mu L, with the detection limit of 0.012 pg/mu L. 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. (C) 2011 Elsevier Inc. All rights reserved.