MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, South China Normal University , Guangzhou 510631, China.
Anal Chem. 2016 Oct 18;88(20):10191-10197. doi: 10.1021/acs.analchem.6b02772. Epub 2016 Sep 26.
Genetic analysis is of great importance for the detection of pathogenic bacteria. Bacterial identification must become simpler, less expensive, and more rapid than the traditional methods. In this study, a low-cost, label-free, and wireless paper-based bipolar electrode electrochemiluminescence (pBPE-ECL) analysis system was constructed for the rapid and sensitive genetic detection of pathogenic bacteria. Wax-screen printing was used to form hydrophilic channels on filter paper, and a carbon ink-based bipolar electrode and driving electrodes were screen-printed into the channels. The "light-switch" molecule [Ru(phen)dppz] (phen = 1,10-phenanthroline; dppz = dipyridophenazine) was used to intercalate into the base pairs of the double-stranded DNA PCR amplification products, and the complexs were then applied to the paper-based bipolar electrode to perform the ECL assays; the ECL of [Ru(phen)dppz] is quenched in aqueous solution, but this molecule displays intense ECL when intercalated into double-stranded DNA. Under optimized experimental conditions, as little as 10 copies/μL of the genomic DNA of Listeria monocytogenes could be detected. Additionally, the system could also specifically distinguish Listeria monocytogenes from Salmonella, Escherichia coli O157:H7, and Staphylococcus aureus. This label-free, simple, and rapid method has potential in point-of-care applications for pathogen detection.
遗传分析对于致病菌的检测非常重要。细菌鉴定必须比传统方法更简单、更廉价、更快速。在这项研究中，构建了一种低成本、无标记、无线的纸基双极电极电化学发光（pBPE-ECL）分析系统，用于快速灵敏地遗传检测致病菌。蜡印版用于在滤纸上形成亲水通道，并用碳墨基双极电极和驱动电极印刷到通道中。将“光开关”分子 [Ru(phen)dppz]（phen = 1,10-菲咯啉；dppz = 二吡咯并[3,2-a:2',3'-c]吩嗪）用于嵌入双链 DNA PCR 扩增产物的碱基对中，然后将复合物应用于纸基双极电极上进行 ECL 测定；[Ru(phen)dppz]的 ECL 在水溶液中被猝灭，但当插入双链 DNA 时，该分子会显示出强烈的 ECL。在优化的实验条件下，仅需 10 拷贝/μL 的单核细胞增生李斯特菌基因组 DNA 即可检测到。此外，该系统还可以特异性地区分单核细胞增生李斯特菌与沙门氏菌、大肠杆菌 O157:H7 和金黄色葡萄球菌。这种无标记、简单、快速的方法在病原体检测的即时护理应用中具有潜力。
