Zhang Ze, Yu Hong-Wei, Wan Guang-Cai, Jiang Jing-Hui, Wang Na, Liu Zhi-Yong, Chang Dong, Pan Hong-Zhi
Fudan University Pudong Medical Center, Shanghai Pudong Hospital, Department of Clinical Laboratory, 2800 Gongwei Rd, Huinan Town, Pudong, Shanghai 201399, China.
The First Affiliated Hospital of the Harbin Medical University, No. 23 Youzheng St, Harbin, Heilongjiang Province 150001, China.
J AOAC Int. 2017 Mar 1;100(2):548-552. doi: 10.5740/jaoacint.16-0251. Epub 2016 Nov 30.
A label-free DNA hybridization electrochemical sensor for the detection of Klebsiella pneumoniae was developed, which could be helpful in the diagnosis of bacterial infections. Indole-5-carboxylic acid (ICA) and graphene oxide (GO) were electrodeposited on a glassy carbon electrode, and the resulting reduced GO (rGO)-ICA hybrid film served as a platform for immobilizing oligonucleotides on a single-stranded DNA (ssDNA) sequence. The conditions were optimized, with excellent electrochemical performance. A significant change was observed after hybridization of ssDNA with the target probe under optimum conditions. Hybridization with complementary, noncomplementary, one-base mismatched, and three-base mismatched DNA targets was studied effectively by differential pulse voltammetry. The proposed strategy could detect target DNA down to 3 × 10-11 M, with a linear range from 1 × 10-6 M to 1 × 10-10 M, showing high sensitivity. This electrochemical method is simple, free from indicator, and shows good selectivity. Hence, electrochemical biosensors are successfully demonstrated for the detection of K. pneumoniae.
开发了一种用于检测肺炎克雷伯菌的无标记DNA杂交电化学传感器，这可能有助于细菌感染的诊断。将吲哚-5-羧酸（ICA）和氧化石墨烯（GO）电沉积在玻碳电极上，所得的还原氧化石墨烯（rGO）-ICA混合膜用作在单链DNA（ssDNA）序列上固定寡核苷酸的平台。对条件进行了优化，具有优异的电化学性能。在最佳条件下，ssDNA与靶探针杂交后观察到显著变化。通过差分脉冲伏安法有效地研究了与互补、非互补、单碱基错配和三碱基错配DNA靶标的杂交。所提出的策略能够检测低至3×10-11 M的靶DNA，线性范围为1×10-6 M至1×10-10 M，显示出高灵敏度。这种电化学方法简单、无需指示剂，且具有良好的选择性。因此，成功地证明了电化学生物传感器可用于检测肺炎克雷伯菌。
