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Rapid detection of foodborne pathogen Listeria monocytogenes by strand exchange amplification

Zhang, Meiling, Wang, Xiudan, Han, Lingzhi, Niu, Shuyan, Shi, Chao, Ma, Cuiping
Analytical biochemistry 2018 v.545 pp. 38-42
DNA, Listeria monocytogenes, bacteria, equipment, food pathogens, gene amplification, genes, point-of-care testing, rapid methods, reverse transcription, ribosomal RNA
A strand exchange amplification (SEA) method to detect foodborne pathogen Listeria monocytogenes was developed. SEA is a novel nucleic acid amplification method that only requires one pair of primers. The specie-specific primers were designed by targeting the 16S rRNA gene and the amplification reaction was performed as short as 60 min at 61 °C. Notably, SEA method could not only detect genomic DNA but also detect RNA by one step without requiring extra reverse transcription. The result could be visualized by naked eyes so that water bath pot would be the only equipment needed. Moreover, culture fluids and bacteria colony could be successfully detected without any pretreatment and the method displayed good specificity and strong anti-jamming capacity. These features greatly simplified the operating procedure and made SEA method be potential for developing point-of-care testing (POCT) devices to detect viable L. monocytogenes.