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Paper-Based Bipolar Electrode Electrochemiluminescence Switch for Label-Free and Sensitive Genetic Detection of Pathogenic Bacteria
- Liu Hongxing, Zhou Xiaoming, Liu Weipeng, Yang Xiaoke, Xing Da
- Analytical chemistry 2016 v.88 no.20 pp. 10191-10197
- DNA, Escherichia coli O157, Listeria monocytogenes, Salmonella, Staphylococcus aureus, aqueous solutions, bacteria, carbon, electrochemiluminescence, electrodes, hydrophilicity, microbial detection, polymerase chain reaction, rapid methods
- 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)₂dppz]²⁺ (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)₂dppz]²⁺ 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.