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用于检测花椰菜花叶病毒35S基因序列的电化学生物传感器,该传感器使用硫化铅纳米颗粒作为寡核苷酸标记物。

Electrochemical biosensor for the detection of cauliflower mosaic virus 35 S gene sequences using lead sulfide nanoparticles as oligonucleotide labels.

作者信息

Sun Wei, Zhong Jianghua, Qin Peng, Jiao Kui

机构信息

College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, Shandong, China.

出版信息

Anal Biochem. 2008 Jun 15;377(2):115-9. doi: 10.1016/j.ab.2008.03.027. Epub 2008 Mar 22.

Abstract

Lead sulfide (PbS) nanoparticles were synthesized in aqueous solution and used as oligonucleotide labels for electrochemical detection of the 35 S promoter from cauliflower mosaic virus (CaMV) sequence. The PbS nanoparticles were modified with mercaptoacetic acid and could easily be linked with CaMV 35 S oligonucleotide probe. Target DNA sequences were covalently linked on a mercaptoacetic acid self-assembled gold electrode, and DNA hybridization of target DNA with probe DNA was completed on the electrode surface. PbS nanoparticles anchored on the hybrids were dissolved in the solution by oxidation of HNO3 and detected using a sensitive differential pulse anodic stripping voltammetric method. The detection results can be used to monitor the hybridization reaction. The CaMV 35 S target sequence was satisfactorily detected with the detection limit as 4.38 x 10(-12)mol/L (3sigma). The established method extends nanoparticle-labeled electrochemical DNA analysis to specific sequences from genetically modified organisms with higher sensitivity and selectivity.

摘要

硫化铅(PbS)纳米颗粒在水溶液中合成,并用作寡核苷酸标记物,用于电化学生物检测花椰菜花叶病毒(CaMV)序列中的35S启动子。巯基乙酸修饰的PbS纳米颗粒能够与CaMV 35S寡核苷酸探针轻松连接。目标DNA序列共价连接在巯基乙酸自组装金电极上,目标DNA与探针DNA的杂交在电极表面完成。通过HNO3氧化溶解固定在杂交体上的PbS纳米颗粒,并采用灵敏的差分脉冲阳极溶出伏安法进行检测。检测结果可用于监测杂交反应。以4.38×10(-12)mol/L(3σ)的检测限令人满意地检测到了CaMV 35S目标序列。所建立的方法将纳米颗粒标记的电化学DNA分析扩展到了对转基因生物特定序列的检测,具有更高的灵敏度和选择性。

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