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Development of DNAzyme-based PCR signal cascade amplification for visual detection of Listeria monocytogenes in food

Liu, Zhanmin, Yao, Chenhui, Yang, Cuiyun, Wang, Yanming, Wan, Sibao, Huang, Junyi
Analytical biochemistry 2018 v.553 pp. 7-11
DNA, Listeria monocytogenes, bacterial contamination, colorimetry, food contamination, food pathogens, microbial detection, oligonucleotides, polymerase chain reaction, pork, rapid methods
Listeria monocytogenes is an important foodborne pathogen, and it can cause severe diseases. Rapid detection of L. monocytogenes is crucial to control this pathogen. A simple and robust strategy based on the cascade of PCR and G-quadruplex DNAzyme catalyzed reaction was used to detect L. monocytogenes. In the presence of hemin and the aptamer formed during PCR, the catalytic horseradish peroxidase-mimicking G-quadruplex DNAzymes allow the colorimetric responses of target DNA from L. monocytogenes. This assay can detect genomic DNA of L. monocytogenes specifically with as low as 50 pg/reaction with the naked eye. Through 20 pork samples assay, visual detection assay had the same results as conventional detection methods, and had a good performance. This is a powerful demonstration of the ability of G-quadruplex DNAzyme to be used for PCR-based assay with significant advantages of high sensitivity, low cost and simple manipulation over existing approaches and offers the opportunity for application in pathogen detection.