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基于纳米金修饰聚邻苯二胺黑 T 膜的电化学生物传感器用于发夹型锁核酸探针检测 BCR/ABL 融合基因

Electrochemical biosensor based on nanogold-modified poly-eriochrome black T film for BCR/ABL fusion gene assay by using hairpin LNA probe.

机构信息

Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou 350004, China.

出版信息

Talanta. 2010 Mar 15;80(5):2113-9. doi: 10.1016/j.talanta.2009.11.017. Epub 2009 Nov 13.

Abstract

A novel electrochemical biosensor is described for detection of breakpoint cluster region gene and a cellular abl (BCR/ABL) fusion gene in chronic myelogenous leukemia (CML) by using thiolated-hairpin locked nucleic acids (LNA) as the capture probe. The hairpin LNA probe was immobilized on the nanogold (NG)/poly-eriochrome black T (EBT) film-modified glassy carbon electrode (GCE). The immobilized LNA probe could selectively hybridize with its target DNA on LNA/NG/EBT/GCE surface. The immobilization and hybridization of the LNA probe were characterized with cyclic voltammetry and electrochemical impedance spectroscopy. The hybridization of the immobilized LNA probe with the target DNA was detected by differential pulse voltammetry with the electroactive methylene blue as an indicator. The results indicated this new method has excellent specificity for single-base mismatch and complementary after hybridization, and a high sensitivity. This novel electrochemical biosensor has been used for assay of PCR real sample with satisfactory result.

摘要

一种新型电化学生物传感器,用于检测慢性髓性白血病(CML)中的断裂点簇区基因和细胞 abl(BCR/ABL)融合基因,采用硫代发夹锁核酸(LNA)作为捕获探针。发夹 LNA 探针固定在纳米金(NG)/聚邻二氮菲黑 T(EBT)修饰的玻碳电极(GCE)上。固定的 LNA 探针可以在 LNA/NG/EBT/GCE 表面上与其靶 DNA 选择性杂交。通过循环伏安法和电化学阻抗谱对 LNA 探针的固定和杂交进行了表征。通过差分脉冲伏安法,用电活性亚甲蓝作为指示剂,检测固定化 LNA 探针与靶 DNA 的杂交情况。结果表明,该新方法对杂交后的单碱基错配和互补具有优异的特异性和高灵敏度。该新型电化学生物传感器已用于聚合酶链反应实际样品的测定,结果令人满意。

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