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

Author:
Liu, Zhanmin, Yao, Chenhui, Yang, Cuiyun, Wang, Yanming, Wan, Sibao, Huang, Junyi
Source:
Analytical biochemistry 2018 v.553 pp. 7-11
ISSN:
0003-2697
Subject:
DNA, Listeria monocytogenes, bacterial contamination, colorimetry, food contamination, food pathogens, microbial detection, oligonucleotides, polymerase chain reaction, pork, rapid methods
Abstract:
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.
Agid:
5969438